Документ взят из кэша поисковой машины. Адрес оригинального документа : http://biochem.bio.msu.ru/assets/docs/diploma/2008/Gluhova.pdf
Дата изменения: Sun Oct 23 15:45:18 2011
Дата индексирования: Mon Oct 1 20:16:17 2012
Кодировка:

..

:

« Hsp20 Hsp27 »

: . . 5 , : ..

, 2008

............................................................................................................... 2 ................................................................................................ 4 ................................................................................................................... 5 .................................................................................................... 6
1. 2. 3. 4. 5. 6. 7. . ......................................................................6 Hsp. ............................................................8 . ............................................10 .....................................................13 . ...............................18 . ......................................20 Hsp20 Hsp27. ..........................................................21

:........................................................................................................ 24 ............................................................................................ 25
1. ....................................................................................25 1.1 1.2 Hsp20Cys ............................................................................................... 25 Hsp27 . ..................................................... 27

2. - Hsp20 Hsp27 ........................................................................................................................................28 2.1 2.2 ............... 28 ................... 29

2.3 Hsp20 Hsp27 . ............................................................................................................. 30 3. Hsp20 Hsp27 ........................30 3.1 - Hsp20 Hsp27. .................................................................................................................... 30 3.2 Hsp20 ............................................................................................ 31 4. ...................................................................................................33 4.1 4.2 4.3 1. ..... 33 SDS. ......................................................... 34 .......................................................................... 34

..................................................................................... 35
- .................................35 1.1 ............ 35 2

1.2 Hsp20 Hsp2740 1.3 Hsp20 Hsp27 ....................... 45 1.4 , «» Hsp20 Hsp27 ............................................................. 49 2. ...............................................56 2.1 2.2 ....................................................................................... 56 Hsp20 .............. 61

............................................................................................................ 70 : .................................................................................................................. 72 ............................................................................................... 73

3

G FRET HEPES Hsp Hsp20cys Hsp27_3D SDS G N,N,N',N'- () (fluorescence resonance en ergy transfer) (4-(2-)-1- (heat shock protein) Hsp20 C46S, A114C Hsp27 c S15D, S78D S82D , - ( mitogen-activated protein kinase activated protein kinase)

4

(heat shock proteins, Hsp) -, , . , . ATP (Hsp60, Hsp70 .), . ( Hsp, small Hsp, sHsp). , . Hsp -, . - , P- . . Hsp , 13 43 . 300-800 . Hsp , . . 10 , Hsp. , , . Hsp20 Hsp27 , , , . , 1% , , Hsp20 Hsp27.

5

1. .
Hsp - N , - , - , (. 1.1). 80-100 , -. (. 1.1). - , , , (van Montfort, et al., 2001). , Hsp . (Kim, et al., 1998). Tsp36 Taenia saginata. - , , , N- Hsp -

. Tsp36 , Hsp - (Stamler, et al., 2005).

N

WDPF

-

C

. 1.1. Hsp16,9 . . . Hsp16,9 . . . (van Montfort, et al., 2001) . 6

-

,

,

, 15-30 . . , Hsp - (Lindner, et al., 2000). Hsp16,9 , - , (van Montfort, et al., 2001) N- Hsp . Hsp . Hsp N- WDPF (. 1.1), . N- - - . , , (Narberhaus, 2002). , N- , Hsp . , 9 24 (Narberhaus, 2002), Hsp (Bukach, et al., 2004; Kim, et al., 2004). Hsp , . . , Hsp (Sobott, et al., 2002), , - -, , (Haley, et al., 1998; Horwitz, et al., 2004) . : Hsp16,5 Methanococcus jannaschii (Kim, et al., 1998) Hsp16,9 (van Montfort, et al., 2001). , Hsp16,9 , , 7

Hsp16,9 (Sobott, et al., 2002). 6 . . , Hsp16,9 (. 1.1). , - , . Hsp16,5 Methanococcus jannaschii Hsp16,9 23%. , , . Hsp16,9, Hsp16,5 Methanococcus jannaschii 24 (Kim, et al., 1998; van Montfort, et al., 2001). , ATP- , , , GroEL. , B- . , , - B- (Haley, et al., 1998). Hsp16,5 Methanococcus jannaschii , , N- . , Thr33, , , (Kim, et al., 1998).

2. Hsp.
, , . Hsp - . , - , B- - 560 , , , (Horwitz, et al., 2004). Hsp Bradyrhizobium japonicum 400 500 (Studer and Narberhaus, 2000). 8

, Hsp (Ehrnsperger, et al., 1999; van de Klundert, et al., 1998). , Hsp25 , Hsp27 , , . Hsp25 . , , (Dudich, et al., 1995; Ehrnsperger, et al., 1999). Hsp20 (43 ) 470 , (van de Klundert, et al., 1998). , Hsp (Bova, et al., 1997; Friedrich, et al., 2004; Sun, et al., 1998). , Hsp (Sobott, et al., 2002; van Montfort, et al., 2001). , . , (Bova, et al., 1997), (Bova, et al., 2002; Friedrich, et al., 2004). , - , , 37°, Hsp16,5 Methanococcus jannaschii ( , 94°) 50° , 72° (Bova, et al., 2002). Hsp16,5 : 30 300 , 5 20% (Bova, et al., 2002). FRET (fluorescence resonance energy transfer), . , . , , , . pH, Ca2+

9

- (Bova, et al., 1997). Hsp . , Hsp25 75° 1,3 , . , (Ehrnsperger, et al., 1999). Hsp27 30 40° , 10° (Lelj-Garolla and Mauk, 2006). Hsp26, Saccharomyces cerevisiae, 24 , , (Haslbeck, et al., 1999). , Hsp . N- (Lambert, et al., 1999; Rogalla, et al., 1999) ( 4 « »). , , . , . - , Hsp. , , , , .

3. .
, , . Hsp , (Lindner, et al., 1997). - ( , pH .). , - (Lee, et al., 1997). (Narberhaus, 2002). 10

Hsp (. 1.2). , (Hsp60 Hsp70) ATP- .

. 1.2. (, et al., 2003). , ATP . , , , Hsp25 , , , . , , . , Hsp ( 62°). Hsp27 , (85°) (Pivovarova, et al., 2005). Hsp (Ehrnsperger, et al., 1997). , , ( .) - , ATP (Biswas and Das, 2007).

11

, Hsp (Ehrnsperger, et al., 1997; Lee, et al., 1997; Lelj-Garolla and Mauk, 2006). Hsp , , , Hsp70 - ATP. , (Friedrich, et al., 2004). . , (2,6 ) B (3 ), - , . (40 ) (Bova, et al., 1997). Hsp . , . Hsp . , Hsp25 Hsp25, Hsp27 (Pivovarova, et al., 2007; Stromer, et al., 2003). . A-, B, 4 37° . 59°, B- , 37°, A- (van Boekel, et al., 1999). - Hsp. , . Hsp , . , Hsp - , .

12

4.
, . Hsp . Hsp . , Hsp20 (Beall, et al., 1999; Beall, et al., 1997), B Hsp27 (Lee, et al., 2005) MAP- (Kato, et al., 1998; Larsen, et al., 1997). . , (Lambert, et al., 1999; Rogalla, et al., 1999). . (Brophy, et al., 1999a; Lambert, et al., 1999; Rogalla, et al., 1999; Theriault, et al., 2004). (Ahmad, et al., 2008). , . Hsp . , . , B- , , Hsp27 (Ito, et al., 1997; Lambert, et al., 1999; Shi, et al., 2008). , Hsp . in vitro , , ,

13

. , Hsp20 (S16D), (Bukach, et al., 2004). Hsp27 . , (Rogalla, et al., 1999). Hsp27 3D (Ser15,78,82D) F-. (Pivovarova, et al., 2005). 3D B- (S19,45,59D) (Ahmad, et al., 2008). , in vivo Hsp27, . . , Hsp27 - Ser15 Ser90. , , (Theriault, et al., 2004). , . , , Hsp27 3 (S15,78,82D) , S15D S78D,S82D(Rogalla, et al., 1999).

Hsp20
N

Ser16
, G

120 137

C

Hsp27
Ser15 Ser78 Ser82

N
MAPKAP MAPKAP MAPKAP, Akt (B), D,

C

. 1.3. Hsp20 Hsp27 . 14

Hsp20 Hsp27, . Hsp20 . G Hsp20 Ser16 ( Hsp20 , , ), ( Hsp20 ), Ser16, (Beall, et al., 1999; Beall, et al., 1997; Brophy, et al., 1999a). Hsp20 , , 120-137 (.1.3) (Beall, et al., 1999). , Hsp20

. G . (Beall, et al., 1997; Brophy, et al., 1999a). , Hsp20 , , ( G) Hsp20 (Brophy, et al., 1999a; Woodrum, et al., 1999). Hsp27 MAPKAP 2 . Hsp27 Ser15, Ser78 Ser82 (Larsen, et al., 1997; Rogalla, et al., 1999). MAPKAP 2 p38 , . , (Knock, et al., 2005; Larsen, et al., 1997). (Shi, et al., al., 1997) Hsp27 2008).

Hsp27 MAPKAP 2 - (Larsen, et

15

Hsp27 , Akt ( B), D (.1.3) (Doppler, et al., 2005; Meier, et al., 2001; Nakajima, et al., 2005; Rane, et al., 2003; Tanabe, et al., 2008). Hsp27 p38 MAP-/Akt (Brophy, et al., 1998; Nakajima, et al., 2005). Hsp25, Hsp27 (Zoubeidi, et al., 2007). Hsp25 , , (Lee, et al., 2005). , Hsp20 Hsp27 . . , Hsp20 , Hsp27, , . , , Hsp20 Hsp27. , Hsp27, Ser15, 78 82, (, ) , . Hsp20 (McLemore, et al., 2005). , , p38 , Hsp27 MAPKAP2 , Hsp20, , (Knoepp, et al., 2000). Hsp27 , Ser78 Ser82. Hsp20, (Fuchs, et al., 2000). Hsp20 Hsp27

16

, - . Hsp20 Hsp27. Hsp25 Hsp20 . , Hsp25 , , , Hsp27 (Benndorf, et al., 1994; Ibitayo, et al., 1999; Wieske, et al., 2001). , Hsp20 (Brophy, et al., 1999b). , - Hsp (Bukach, et al., 2005; Panasenko, et al., 2003). Hsp20 14-3-3. , 14-3-3 , (Birkenfeld, et al., 2003). Hsp20 14-3-3. Hsp20 , 14-3-3 (Chernik, et al., 2007). , 14-3-3, , . , Hsp20 Hsp27 , , . , Hsp in vivo, , , . in vitro . , Hsp27 Hsp20 . Hsp27, MAPKAP 2 , Hsp20,

17

Hsp27 Hsp20. Hsp . , Hsp20 Hsp27 , . Hsp , , - , .

5.

.
, , , . Hsp. , Bradyrhizobium japonicum 10 Hsp, (Studer and Narberhaus, 2000). , (Studer and Narberhaus, 2000). , . , , Hsp (Ganea, 2001). Hsp -, . , - - - B- . , , Hsp. Hsp Bradyrhizobium japonicum Hsp, 18

, , (Studer and Narberhaus, 2000). Hsp , (Fontaine, et al., 2005; Sugiyama, et al., 2000; Sun, et al., 2004). , , Hsp18,1 Hsp16,5 IbpB E. coli Hsp Bradyrhizobium japonicum (Sobott, et al., 2002; Studer and Narberhaus, 2000).

. 1.4. Hsp (, 2006). 10 , Hsp, (. 1.4). -, HspB9 HspB10 ( , ), (MKBP, HspB3 HspB7), Hsp , (Taylor and Benjamin, 2005). . HspB3, MKBP, Hsp27, Hsp20 B-. , Hsp: Hsp20 Hsp27 B- MKBP HspB3. (Sugiyama, et al., 2000). Hsp.

19

6. .
Hsp . , (Sun, et al., 1998). Hsp. , - B-. - B-, 3:1 (Sun and Liang, 1998). Hsp . , Hsp18,1 Hsp16,9 , (Sobott, et al., 2002). -, . , - B . , - pH, . B-, , - (van Boekel, et al., 1999). , . - , 3:1 ( ) (Sun and Liang, 1998). Hsp27 -, Hsp, . 65° Hsp27 , - . , . , Hsp27 -

20

(Fu and Liang, 2003). Hsp27 (Zantema, et al., 1992). . . , Hsp27 - , . -. , , , (Zantema, et al., 1992). , . - B- (Datta and Rao, 2000). . , Hsp Bradyrhizobium japonicum (Studer and Narberhaus, 2000). , , Hsp, . - , . , , Hsp .

7. Hsp20 Hsp27.
1994 . Hsp20 . Hsp20 Hsp27 -, 7 (Kato, et al., 1994). Hsp20 Hsp27 - (Sugiyama, et al., 2000; Sun, et al., 2006). - , Hsp20 Hsp27 , (Sugiyama, et al., 2000). 5 « », Hsp20 Hsp27 21

, ( 1.1) (Kato, et al., 1994; Kato, et al., 1992). Hsp20 Hsp27. 1.1. Hsp20 Hsp27 (Kato, et al., 1994; Kato, et al., 1992). Hsp20, / 12800±2260 ( ) 2050±320 () 281±43 ( ) 1190±180 417±82 191±24 1790±360 24,6±1,6 43,5±7,8 6,4±1,0 3,5±0,3 14,1±4 3,1±0,2 Hsp27, / 2290±320 1710 1170±330 1210±560 971±81 595±303 135±37 246±70 123±70 138 171±17

, Hsp20 Hsp27. - . , ... .. , Hsp20 Hsp27 , - Hsp20 Hsp27 (Bukach, et al., 2004). . , Hsp27 , Hsp20 . , (Fuchs, et al., 2000). , , , ,

22

, . , , Hsp20 Hsp27. . - Hsp20 Hsp27 .

23

:
:
-

Hsp20 Hsp27 .

:
1. 2. 3. 4. 5. Hsp20 Hsp27. Hsp20 Hsp27 . Hsp20 Hsp27 . - Hsp20 Hsp27 -. Hsp20 Hsp27wt , (S15,78,82D).

24

1.
... .. - , , ... .. Hsp20wt.

1.1

Hsp20Cys
Hsp20Cys E. coli. E.coli BL21(DE3), pET23b,

Hsp20Cys, 25 LB, 0,1 / 30° 15-16 . 2 LB, 0,1 / 8-9 37°, 15 30°. (15 5000 g), 20 (50 -HCl, 8,0, 0,1 NaCl, 1 , 14 , 0,5 ) -20°. Hsp20Cys. Hsp20Cys , Hsp20 (Bukach, et al., 2004) . E. coli c 1 , 10 (50 -HCl, 8,0, 0,1 NaCl, 1 , 14 , 0,5 ), 100 (50 /) 30 4°. 35 MgCl2 (2,5 M) 50 I (5 /) 15 . , 20 22000g. , 30 , . , , 30% 1 4°.

25

(20 22000g) , 32 (20 -, 7,6, 10 NaCl, 14 , 0,1 , 0,1 ), 2 . (1 100000g). .

Hsp20Cys. Q-Sepharose HiTrap (V=5 ). , 4 (20 ) (20 -, pH 7,6, 10 NaCl, 14 , 0,1 , 0,1 ). NaCl (00,35 ) . . , Hsp20Cys . , , 0,2 , 8,0. Phenyl-Sepharose Hi-Trap (5 ). 4 (20 -, 8,0, 14 ), 0,2 (NH4)2SO4, . (NH4)2SO4 0,20,01 ; (NH4)2SO4 0,010 , 10 . , , Hsp20Cys, SDS-. , ,

. 20°. 320 1 . coli.

26

1.2

Hsp27 .
Hsp27 ,

(, 2006) . Hsp27. E. coli c 2 , 5 (50 -HCl, 8,0, 0,1 NaCl, 1 , 14 , 0,5 ), 200 (50 /) 30 . 32 MgCl2 (2,5 M) 100 I (5 /) 15 . , 25 23700g. , 30 , . 30% 1 . , 10 (20 -, 7,6, 10 NaCl, 14 , 0,1 , 0,1 ), . (1 100000g). 30% 50%, 1 4°. 10 . (1 100000g). . Hsp27. Q-Sepharose Hi Trap (V=5 ). , 4 (20 ) 1 (20 -, pH 8,0, 10 NaCl, 14 , 0,1 , 0,1 ). NaCl (00,35 ) 1. . ,

27

Hsp27,

(

«»)

Q-Sepharose . - Superdex 200 HR 26/60. , 0,15 NaCl. Hsp27 . , Hsp27, , . 20°.

2.

-

Hsp20 Hsp27
2.1
. , Hsp20 Hsp27 . , Hsp20 (105 ) Hsp27 (105 ) (20 -, pH 7,6, 10 NaCl, 0,1 , 0,1 , 14 ), 15 4° 15 , 30 , 1 3 37° 45°. (0,25 HCl, 6,8, 20% ) (« », 4.1). Hsp20 Hsp27 , Hsp20 (2 /) Hsp27 (2 /) , 37° 30 1 . 4° 15 . (0,25 -HCl, 6,8, 20% ) . , 37° 1 30 .

28

-. -. 100 , Hsp20 (100 ) Hsp27 (100 ), 1 3 37° 1 45°. 10 (200 -, 7,6, 100 NaCl, 140 , 1 , 1 ) Superdex 200 HR10/30, , 150 NaCl. 100 . , . Image Quant, Hsp27/Hsp20 .

2.2
Hsp20 Hsp27 -. . (V=100 ), Hsp20 Hsp27, ( 30 150 ), 3 37°. , 10 (200 -, 7,6, 100 NaCl, 140 , 1 , 1 ) Superdex 200 HR 10/30, , 150 NaCl. 100 . , Hsp20 Hsp27 3 37° ( 150 ). , , , . , . Image Quant, Hsp27/Hsp20 .

29

Hsp20, Hsp27 . Hsp20 Hsp27 (44 ) 1 45°, . , Hsp20 (88 ) Hsp27 (88 ) 50 , pH 7,5, 150 NaCl, 2 . 20 44 ( Hsp20 Hsp27). Beckman E, 0,6°, 8000 / ( 7,5 ). 292 . SEDFIT.

2.3 Hsp20 Hsp27 .
Hsp20 Hsp27 , , 210 , Hsp20/Hsp27 5/1, 3/1, 2/1, 1/1, 1/2, 1/3, 1/5. 3 37°, . , Hsp20 Hsp27 , - (« », 2.1 2.2).

3.

Hsp20

Hsp27
3.1 - Hsp20 Hsp27.
- . - Hsp20 Hsp27 . -. 30

, . (Farahbakhsh, et al., 1995) . (7,2 /), 2,5% , (300 ) , 0,24 /. 10 37° 20 . (50 , 7,5, 50 NaCl,) 37°. Hsp20 Hsp27 1,5 12 , 0,75 . Hsp20 Hsp27 1 45° 3 37°. Hsp20 Hsp27, 3 37° 1 45°. 340 Ultraspec 3100 pro. - . 5 (30 HEPES, 1 MgCl2, pH 7,3). F- (1 /), 58,5° ( ). 340 Cary Eclipse (Varian). . Hsp20, Hsp27 1,5 , 0,75 58,5 °. . .. .. . .. .

3.2 Hsp20

Hsp20 Hsp27 . Hsp20 () Hsp20 (110

31

) (20 -, 7,6, 10 NaCl, 14 , 0,1 , 0,1 ) ATP (0,4 ), ( ) (2*10-3 /), ( 400 150 ) -32P ATP (25 HEPES-NaOH, 7,5, 25 Na-, 20 MgCl2, 2 ), 30° 30 , 1 2 . Hsp20 Hsp20 Hsp20 (110 ) Hsp27 (110 ) 1 37° 1 45° . , . ATP . ( 4.1) ( 4.2). . Hsp20 Hsp27 Ser (Hsp27_3D) . Hsp20 Hsp27_3D ( 110 ) 45° . , , . Hsp20. , Hsp20 (50 ) Hsp27 (0 - 200 ), 1 45° Hsp20 Hsp27 (1/0; 2/1; 1/1; 1/2 1/4). , ATP ( 200 ), -32P ATP . 90 30°, (20 ) Whatmann 3MM. 3 20 10% , 5 Na2HPO4 5 Na2P2O7, 3 96% . ( POPOP PPO 32

) RackBeta 1214 (LKB) . Hsp20 1 Hsp20 1 ATP . Hsp20 Hsp27wt Hsp27_3D, Hsp27wt 1 45°. Hsp27 Hsp20 . .. .

4.

4.1
(6 15%) . 2.1. 2.1. 30% , 0,8% , , , 10 % , , 2 4% 0,27 0,5 (0,5 -HCl, 6,8) 1,23 10 2 4 6% 0,8 1 (1,5 -HCl, 8,8) 2,2 10 2 4 15% 2 1 (1,5 -HCl, 8,8) 1 10 1

(0,25 -HCl, 6,8, 40% ). : (0,025 -HCl, 8,6), (0,192 -, 8,6). 10 / 3 . 10%

33

, 0,2% R-250, 10% 30% . . - (440 ), (66 128 ) (43 ).

4.2

SDS.
: 15% ,

0,4% , ј (1,5 -HCl, 8,8, 0,4% SDS), 0,05% . 0,1%. : 6% , 0,16% , ј (0,5 HCl, 6,8, 0,4% SDS), 0,05% . 0,1%. 1/3 (0,25 -HCl, 6,8, 8% SDS, 40% , 0,004% , 5% ). : - 0,192 -, 8,6, 0,1% SDS; 0,025 -HCl, 8,6. 10/ 20 / . 10% , 0,2% R-250, 10% 30% . .

4.3

.

, 2.2.

2.2. . 280 0,556 1,779 0,955

Hsp20 Hsp27

Swiss-Prot: 14558 Swiss-Prot: 04792 Swiss-Prot: 01317 34

1. -

1.1

, . , (Bova, et al., 1997). Hsp , .

- (Sun, et al., 1998), . Hsp20 Hsp27 . , (Kato, et al., 1994; Kato, et al., 1992). , 37°. , Hsp27 , (Sugiyama, et al., 2000), , . , (37°) (45°). Hsp20 Hsp27 -. , (. 3.1), 4° 15 . , Hsp20 Hsp27 (. 3.1, 1).

35

200

150

3, 4 D
280
100

2
50

0 8 10 12 14 16

1
18

,
. 3.1. Hsp20 Hsp27. . . Hsp20 (105 ) Hsp27 (105 ) 15 4є (1), 15 (2), 30 (3) , 1 (4) 3 (5) 45° 15 (6), 30 (7), 1 (8) 3 (9) 37°. Hsp20, Hsp27, - . . - Superdex 200 HR 10/30. , , 150 NaCl, 100 Hsp20 (100 ) Hsp27 (100 ), 4° (1), 1 37° (2), 3 37° (3) 1 45° (4). . 36

37°, , Hsp20 Hsp27 170 260 . 3 (. 3.1, 6-9). 37° , Hsp20 Hsp27. 45°C, Hsp20 Hsp27 15 (. 3.1, 2-5). , , Hsp20 Hsp27, , , 60°, ( ). , , (. 3.1). Hsp20 Hsp27, 4°, 730 50 , Hsp27 Hsp20 (. 3.1, 1). 1 37°C 3 630, 57 120 , , , Hsp27 Hsp20, . Hsp27 (630 kDa) (734 kDa), Hsp20 (50 57 kDa). , - - . C , , (. 3.1), , Hsp27 , , Hsp27, Hsp20. , Hsp20 . , , 37°C Hsp20 Hsp27

37

, , . Hsp20 Hsp27, 3 37°, , 400 140 (. 3.1, 3). , 1 45°C (. 3.1, 4). , , , , . , , 37° 45° Hsp20 Hsp27. 4° , , , Hsp20 Hsp27. Hsp20 Hsp27 37° , (30 1 ), 4° (15 ) , 37° (. 3.2).

. 3.2. Hsp20 Hsp27 4. . Hsp20 Hsp27 ( 2 / ), 15 4° (1), 30 37° (2), 30 37° 4° 15 (3), (4), 1 37° 4° 15 (5), Hsp20 (6) Hsp27 (7). 38

, 4° (. 3.2, 2 3; 4 5). , , . , , 4°, Hsp20 Hsp27 3 37° 15 45°, . . , Hsp (Bova, et al., 1997; Friedrich, et al., 2004; Sun, et al., 1998). (Bova, et al., 1997). , Hsp. Hsp20 Hsp27 , - (Sun, et al., 1998). 45°, , , . Hsp . (, Hsp25 ), , (, Hsp26 ) (Ehrnsperger, et al., 1999; Haslbeck, et al., 1999). Hsp20 Hsp27 4° 45° - . (. 3.1, 2-5 9), - (. 3.1, 3 4) , . , , , , Hsp20 Hsp27 39

, . .

1.2

Hsp20 Hsp27
« », , (Ehrnsperger, et al., 1999; van de Klundert, et al., 1998). , Hsp Hsp20 Hsp27 . , , . Hsp20 Hsp27 (150, 100, 50 30 ) 3 37° . - (. 3.3, ). , Hsp20 Hsp27 150 , , , ( 3.3, ). , (30 ), , (. 3.3, 1), 5 , (. 3.3, 4). , , , Hsp20 Hsp27, . , , .

40

. 3.3. Hsp20 Hsp27. Superdex 200, , 150 NaCl, Hsp20 Hsp27, 3 37°. , 150 ( ) ( ). 1 200 , 150 ; 2 100 , 150 ; 3 100 , 100 ; 4 100 , 50 ; 5 100 , 30 ; 6 Hsp20 Hsp27 200 , 25 . - . 41

, , . , , , , , . , (. 3.3). , , , . 30 (. 3.3, 2) 6 (. 3.3, 5), 420 340 , 175 105 . , , , . , (- ) , , . - , . , , , , . , , , «» , . , , , Hsp20 Hsp27 , 42

, . Hsp, , Hsp25 Hsp20 (Ehrnsperger, et al., 1999; van de Klundert, et al., 1998). , Hsp25 , , , (Dudich, et al., 1995; Ehrnsperger, et al., 1999). , Hsp20 Hsp27 . . , , , -. , , . . SEDFIT, (s) (Schuck, 2000). S , . :

, M , f , - , (, ). . (f/f0), f , f0 , , . s, f/f0 :

43

, M , f /f0 , , (, ), - , Na . . 3.4 (s) Hsp20, Hsp27 .
0.4

2

0.3

3 c(s)
0.2

1

0.1

0.0

4

8

12

16

20

24

, s
. 3.4. Hsp20, Hsp27 . (s). 1 Hsp20 (40 ); 2 - Hsp27 (40 ); 3 - Hsp20 (20 ) Hsp27 (20 ) Hsp20 Hsp27 1 45°C. Hsp20 3,7s 6,2s, , , (. 3.4, 1). Hsp27 , 15s, , , (. 3.4, 2). Hsp20 Hsp27, , , 11s (. 3.4, 3). , , Hsp20, Hsp27

44

, , , - . , (s) SEDFIT f/f0 . Hsp, , , f/f0 . , . - . , , , , Hsp20 Hsp27 . 3 37°, 45°. Hsp20 Hsp27 , . , Hsp20 Hsp27 ( 1 1) .

1.3

Hsp20 Hsp27

, , Hsp20, - Hsp27. - . Hsp20 (Bukach, et al., 2004), Hsp27 , 700 (. 1.1 « »).

45

, . Superdex 200 10 30 Hsp20 Hsp27, (. 3.5 , ). 10 Hsp20 Hsp27 1 37°, , (. 3.5). Hsp27/Hsp20, Hsp20 Hsp27. . 3.5-, ( ) Hsp27/Hsp20 ( ). , Hsp27/Hsp20 , Hsp27, , Hsp20 (. 3.5, . . 3.3). , . Hsp27 Hsp20 ( 42-46), . ( 3 37°), , , , Hsp27 Hsp20 (. 3.5 ). , . , Hsp27 (. 3.5 , 32-36), Hsp27, Hsp20 (. 3.5 , 46-50). , Hsp27 Hsp20 , . , Hsp27/Hsp20 .

46

80 70 60

28 30 32 34 36 38 40 42 44 46 48 50 52 54 56

400

80

28 30 32 34 3 6 3 8 4 0 4 2 44 46 48 50 52 5 4 5 6

20 15 4 3 2 1

Hsp27/Hsp20

350 4 3 2 1 0 18

60

40 30 20 10 0 10 12 14 16

D280, mAU

D280, mAU

50

50 40 30 20 10 0 10 12 14 16

0 18

,

,

250 200

28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 5 4 3 2 1 0 18

Hsp27/Hsp20

D280, mAU

150 100 50 0

10

12

14

16

,

. 3.5. Hsp27/Hsp20 . -. Superdex 200 HR 10/30 Hsp20 Hsp27 1 (), 3 () 37° 100 ( ), 100 , 3 37° 150 , 200 (). Hsp27/Hsp20, SDS . . . , 3 37°, Hsp20 Hsp27 (210 ), Hsp20/Hsp27. 1 5/1, 2 3/1, 3 2/1, 4 1/1, 5 1/2, 6 1/3, 7 1/5, 8 - Hsp20 Hsp27 1/1, Hsp20, Hsp27, - .

Hsp27/Hsp20

70

47

« », (Sobott, et al., 2002). , , Hsp20/Hsp27 1. . (210 ), Hsp27 Hsp20. 3 37° ( 3.5). Hsp20 Hsp27. , Hsp27 Hsp20 (. 3.5, 4). , Hsp. Hsp20 , Hsp20 (. 3.5, 1-3). Hsp27 (. 3.5, 5-7). , , Hsp27/Hsp20 . Hsp , . - B-, , 3:1 (Sun and Liang, 1998). . Hsp18,1 Hsp16,9 , (Sobott, et al., 2002). , in vivo, . , Hsp20 Hsp27 , . , Hsp20 Hsp27 ( 1.1),

48

, Hsp20 Hsp27.

1.4

, «»

Hsp20 Hsp27
Hsp20 Hsp27, , , «» . , , , , Hsp20 Hsp27 . Hsp20 Cys46, N- , Hsp27 - Cys137, - (.3.6). ,

Hsp20wt
N

Cys46

C
Cys137

Hsp27
N

C
Cys46Ser Ala114Cys

Hsp20cys
N

C

. 3.6. Hsp20wt, Hsp27 Hsp20cys . , , Hsp20 Hsp27, , . « », Hsp , « » , , . , ,

49

.

,

, , -. , B- - , N- (Sreelakshmi, et al., 2004). , , Hsp20 Hsp27, . Hsp20 Hsp27 , «» . Hsp20 Hsp27 , «» . Hsp20cys (C46S, A114C) «» , Cys137 Hsp27 (.3.6). , Hsp20cys, Hsp27 Hsp20wt . Hsp20 Hsp27 , Hsp27 Hsp20wt Hsp20cys, , . . (. 3.7) (. 3.7). (. 3.7, «+»). , «» Hsp20 ( C46S, A114C) Hsp27 (. 3.7, «+», 4 5). Hsp20wt , «» (. 3.7, «-», 1), . , (. 3.7, «-», 1). , Hsp20 , . , -,

50

. 3.7. Hsp20 Hsp27 , . , Hsp20wt (1), Hsp20cys (2), Hsp27 (3) Hsp27 Hsp20wt (4) Hsp20cys (5) (+), (-). , . . . .

. 3.8. Hsp20 Hsp27. , Hsp20wt (1), Hsp20cys (2) Hsp27 (3) , 1 45° (+) (-). (-) (+) Hsp27 Hsp20wt (4) Hsp20cys (5). () (), . 51

, . Hsp27 «» 10 40%. Hsp20cys, , Cys137 Hsp27 (. 3.7, «-», 2 3). , Hsp20cys, Hsp27 , (. 3.7, 2, 3 5). Hsp20wt Hsp27, Hsp20wt, , . , , , (. 3.7, «-», 4). Hsp27 Hsp20wt . , . , , «» . Hsp20, Hsp27 , Hsp20wt, Hsp20cys , . - «» Hsp27, , . , Hsp27 , Hsp20 Hsp27. Hsp20cys Hsp27 , «»

(. 3.5, 2, 3 5). , Hsp20cys Hsp27 . , , . , , , . 52

1h 45° . Hsp20wt Hsp27 , ( ). , , Hsp20wt, Hsp27 - . (.3.8). Hsp20wt, Hsp20cys Hsp27 , , . , 1 45°, , . , , . (. 3.8) (. 3.8). , Hsp20 , , Hsp20cys Hsp27 (. 3.8, «-», 2 3). 1 45° Hsp20 Hsp27 (. 3.8, «-», 4 5) Hsp20, . , Hsp20wt, Cys46. , Hsp27 Hsp20wt Hsp20cys, Hsp27 , Hsp20, , . , Hsp20wt Hsp20cys , , , , - Hsp20.

53

, , Hsp20 Hsp27 , Cys46 Hsp20 , Hsp20 . , , , Cys137 Hsp27 ( Cys114 Hsp20) , Hsp27 Hsp20. Hsp20 Hsp27 , , . Hsp20 Cys46 N- . Hsp16.9 Triticum aestivum, , N Hsp (van Montfort, et al., 2001). , Hsp27 (Cys137) - , , ( ) «» Hsp27 , Hsp27. ( ). , Hsp20 , , (.3.9). Hsp27

. 3.9 - Hsp20 Hsp27. , . , Hsp20, , . , Hsp20cys, , 54

Hsp27, . , , , , . , , Hsp (Sobott, et al., 2002; van Montfort, et al., 2001). Hsp20 Hsp20 Hsp27 (« », 1.3), , , 2 .. ( 100 ), 4 , , Hsp20, Hsp27. , , , «» Hsp20 , Hsp20 Hsp27. , Hsp20 . , Hsp27 . , , , Cys137 Hsp27, Hsp20. Hsp20wt, Cys137 Hsp27, Hsp20Cys .

55

2.
2.1

,

, Hsp. « », . (van Boekel, et al., 1999). - (Ganea, 2001). - . - , , , Hsp. Hsp20 Hsp27, . 1 45° 3 37°. , . , , , , 340 (. 3.10, 1). Hsp20 ( 2) Hsp27 ( 3) 1,5 - . Hsp , ( ). . 3.10, Hsp20 Hsp27, , (. 3.10, 2 3). 3 37° 1 45° ( ).

56

, %

100 80 60 40 20 0

1 2 3 4,5,6

0

5

10

15

20

25

30

,
. 3.10. Hsp20, Hsp27 -. (1), Hsp20 (2), Hsp27 (3), Hsp20 Hsp27 (4) (5 6). 3 37° (5) 1 45° (6). 0,24 /, Hsp20 Hsp27 - 1,5 . Hsp20 Hsp27 1,5 ( 0,75 ). D340 , . Hsp20 Hsp27 ( ) . ,

, Hsp20 Hsp27 3 37° 1 45° (. 3.10, 5 6). , Hsp20 Hsp27 (. 3.10, 4, 5 6). . -, , , , , . Hsp20 Hsp27 . 57

( Hsp20 Hsp27) . Hsp . . Hsp20 Hsp27 , , Hsp. .. , - , , , . , Hsp . , . Hsp20, Hsp27 F-. , . , . Hsp . , (Lambert, et al., 1999; Panasenko, et al., 2003), -. (. 3.11). (1°/) , , 340 (. 3.11, 1). Hsp20, Hsp27 ( 1,5 ), 3 37°, (. 3.11, 2, 3 4),. , Hsp20 Hsp27, , 58

, .

600 500 400 300 200 100 0 20

1 2 3,4

1 2 4 3

30

40

50

60

70

80

90 0

10

20

, °

30 ,

40

50

. 3.11. Hsp20, Hsp27 F- -. . F 1 /. . F- 58,5°. 1 , 2 - 4 Hsp20 (2), Hsp27 (3) (4). 3 37°. 0,2 /, Hsp20 Hsp27 - 1,5 () 0,75 (). Hsp20 Hsp27 1,5 () 0,75 ().

. . . 3.11 , Hsp , 58-59°. , 58,5° 1 . 340 (. 3.11). 58,5° Hsp, 1,5 ( Hsp20 Hsp27), ( ), , 0,75 Hsp20 Hsp27, . , Hsp20, Hsp27

59

. . , Hsp20, Hsp27 -, Hsp20 Hsp27, . Hsp20 Hsp27 -, Hsp20 Hsp27 , . . . - B- , Hsp Bradyrhizobium japonicum (Datta and Rao, 2000; Studer and Narberhaus, 2000). - Hsp, . , , . , 1 45 ° . ., Hsp27 75° (Ehrnsperger, et al., 1999). Hsp , . . Hsp , .

60

2.2

Hsp20

« », Hsp20 (Beall, et al., 1997; Brophy, et al., 1999a; Woodrum, et al., 1999). Hsp20 () Ser16 (Beall, et al., 1999). Hsp20, Hsp27. Hsp27 cAMP- , (Ser15,78 82) MAPKAP 2 , (Brophy, et al., 1998; Knoepp, et al., 2000). Hsp20 Hsp27 , . Hsp27 Hsp20 cAMP- ( , ). , Hsp20 Hsp27wt, (. 3.12 ). Hsp20 , -32P ATP. (. 3.12) (. 3.12). , . , Hsp20 (. 3.12, 1 2), , . , Hsp20, , , , . . , . , , Hsp20. 61

, %
60 40

1 2 3 4 5

20

0

0

40

80

120

160

,
. 3.12. Hsp20 Hsp27wt . , Hsp20wt (1), Hsp20 Hsp27wt (2), (3) (4) Hsp20 Hsp27wt (4), 30 - ATP . () (). . ( ) (- ). . Hsp20 . 1 Hsp20, 2-5 Hsp20 Hsp27. Hsp20/Hsp27wt 2/1 (2), 1/1 (3), 1/2 (4), 1/4 (5).

62

( ). , , , . , Hsp20 . Hsp27wt Hsp20 (. 3.12 , 2). Hsp20 (. 3.12, 2). , Hsp20 Hsp27 , Hsp20 (. ). Hsp20 , , Hsp27 Hsp20 (. 3.12, 4). Hsp20 (Hsp20 Hsp27 1 37°) , Hsp20, Hsp27 (. 3.12, 3). Hsp20 Hsp20 Hsp27 (. 3.12). , Hsp20 , . . , Hsp20 , Hsp20. , , Hsp20 Hsp27 . , . , , Hsp20, . Hsp20 , , , , . Hsp20 Hsp27 ( 1.1, . 22). 63

. , Hsp27 , Hsp20/Hsp27 . Hsp20 Hsp27, Hsp20 Hsp27 ( Hsp20 50 , Hsp27 0 200 ). Hsp20 , (. 3.12.). Hsp20, , , , , 100%, (. 3.12, 1). , -32P ATP Hsp20. , Hsp20, Hsp27 Hsp27 (. 3.12, 2-5). Hsp20 Hsp27 Hsp20 (. 3.12, 5). , , Hsp20 . , Hsp27, Hsp20 Hsp27 , Hsp20 , . , , Hsp20/Hsp27. , , , , Hsp27 Hsp20, , (. 3.5). , Hsp20 Hsp27, . , Hsp27, , Hsp20 Hsp27. Hsp20 , Hsp20 Hsp27. 64

Hsp27, Hsp20 . , , Hsp27 Hsp20 . , Hsp20 Hsp27_3D. Hsp27, (Ser15,78 82) MAPKAP 2 (Rogalla, et al., 1999). , Hsp27 ( Hsp27 . Hsp20, Hsp27_3D (. 3.13). Hsp20 - -32P ATP. , (. 3.13 ). Hsp20 Hsp27_3D Hsp27 (. 3.12, 4 . 3.13, 2). Hsp27, , , . . 3.13 , Hsp20 Hsp27_3D . Hsp20, , Hsp27_3D (. 3.13). , Hsp27_3D , Hsp20. , Hsp20 , , Hsp20, Hsp27wt . Hsp27_3D ( ), Hsp20 , . , Hsp20 Hsp27_3D . Hsp20 , . 3D ) Hsp20, ,

65

, %
60 50 40 30 20 10 0

1,2,3, 4,5

0

20

40

60

,
. 3.13. Hsp20 Hsp27_3D . , Hsp20 (1) Hsp20 Hsp27_3D (2), 30 - ATP . () (). . ( ) (- ). . Hsp20 . 1 Hsp20, 2 5 Hsp20 Hsp27_3D. Hsp20/Hsp27_3D 2/1 (2), 1/1 (3), 1/2 (4), 1/4 (5).

66

., , Hsp27wt Hsp20, Hsp27, MAPKAP 2 , (Fuchs, et al., 2000). , . , Hsp27 (Hsp27_3D), , MAPKAP 2 . , Hsp27. , Hsp27 , ,

. , Hsp27 Hsp27wt (Hsp27_3D). . Hsp20 Hsp27wt, . . 3.14 Hsp20 Hsp27wt, Hsp27_3D (. 3.14).

, %

70 60 50 40 30 20 10 0 0 20 40 60 80

3 4 1 2

100

,
. 3.14. Hsp20 Hsp27wt, Hsp27_3D . Hsp20 (1), Hsp20 Hsp27wt (2), Hsp27_3D (3) Hsp27wt Hsp27_3D (4). Hsp20/Hsp27 1/2.

67

Hsp20 : Hsp27wt, Hsp27_3D , Hsp20. , . , , . , Hsp27_3D MAPKAP 2 Hsp27wt. , . , Hsp27 MAPKAP 2 . Hsp27 Hsp20. , Hsp27 ,

(Lambert, et al., 1999; Rogalla, et al., 1999). , Hsp20 Hsp27_3D , Hsp27wt (Bukach, et al., 2004). Hsp20 Hsp27 ( Hsp27wt Hsp27_3D) Hsp27wt Hsp27_3D ( Hsp27). , Hsp27 Hsp20. , Hsp20 , Hsp20 . , Hsp27wt Hsp27_3D . , Hsp20 Hsp27wt Hsp27_3D, , Hsp20, Hsp27wt Hsp27_3D. Hsp20 . , , Hsp27 ., , . Hsp20 Hap27 , 37° , in vivo . , , , 68

. , , Hsp20 Hsp27. , ., , (Fuchs, et al., 2000).

69

, , , Hsp20 Hsp27, , , . , , (Bova, et al., 1997; Sun, et al., 1998). 4° , Hsp, . , (Ehrnsperger, et al., 1999; van de Klundert, et al., 1998). Hsp20 Hsp27 , . Hsp20/Hsp27 , , « » . , Hsp20 Hsp27 - , . Hsp20/Hsp27 , , . , - (Sreelakshmi, et al., 2004). Hsp20 Hsp27, , , Hsp20 Hsp27 . , Hsp20 , Cys46. N- Hsp20,

70

Hsp, , Hsp20 , . , Cys138 Hsp27, , , , . Hsp27 , , Hsp27, , Hsp27 Hsp20. , , Hsp20 Hsp27 . Hsp20 Hsp27, , Hsp20 Hsp27 . Hsp20 Hsp27 , -, . , , , Hsp20 Hsp27. , -. , Hsp20 Hsp27 . , , (Beall, et al., 1997; Brophy, et al., 1999a; Brophy, et al., 1998; Knoepp, et al., 2000). , Hsp20 Hsp27wt Hsp20 . , Hsp20 Hsp27_3D . , Hsp20 , , , .

71

:
1. Hsp20 Hsp27 , - , Hsp20 Hsp27 1. 2. 37° 45° Hsp20 Hsp27, 4° . 3. Hsp20 Hsp27 , «» . Hsp27 . 4. Hsp20 Hsp27 -. , . F-, - Hsp20 Hsp27 . 5. Hsp27 Hsp20

, Hsp27 , (S15,78,82D), .

72

, . . (2006). 20 (Hsp20, HspB6). . , ., , . and , . . (2003). . 43, 59-98. Ahmad, M. F., Raman, B., Ramakrishna, T. and Rao Ch, M. (2008). Effect of phosphorylation on alpha B-crystallin: differences in stability, subunit exchange and chaperone activity of homo and mixed oligomers of alpha B-crystallin and its phosphorylation-mimicking mutant. J Mol Biol 375, 1040-51. Beall, A., Bagwell, D., Woodrum, D., Stoming, T. A., Kato, K., Suzuki, A., Rasmussen, H. and Brophy, C. M. (1999). The small heat shock-related protein, HSP20, is phosphorylated on serine 16 during cyclic nucleotide-dependent relaxation. J Biol Chem 274, 11344-51. Beall, A. C., Kato, K., Goldenring, J. R., Rasmussen, H. and Brophy, C. M. (1997). Cyclic nucleotide-dependent vasorelaxation is associated with the phosphorylation of a small heat shock-related protein. J Biol Chem 272, 11283-7. Benndorf, R., Hayess, K., Ryazantsev, S., Wieske, M., Behlke, J. and Lutsch, G. (1994). Phosphorylation and supramolecular organization of murine small heat shock protein HSP25 abolish its actin polymerization-inhibiting activity. J Biol Chem 269, 20780-4. Birkenfeld, J., Betz, H. and Roth, D. (2003). Identification of cofilin and LIM-domaincontaining protein kinase 1 as novel interaction partners of 14-3-3 zeta. Biochem J 369, 45-54. Biswas, A. and Das, K. P. (2007). alpha-Crystallin Assisted Refolding of Enzyme Substrates: Optimization of External Parameters. Protein J. Bova, M. P., Ding, L. L., Horwitz, J. and Fung, B. K. (1997). Subunit exchange of alphaA-crystallin. J Biol Chem 272, 29511-7. Bova, M. P., Huang, Q., Ding, L. and Horwitz, J. (2002). Subunit exchange, conformational stability, and chaperone-like function of the small heat shock protein 16.5 from Methanococcus jannaschii. J Biol Chem 277, 38468-75. Brophy, C. M., Dickinson, M. and Woodrum, D. (1999a). Phosphorylation of the small heat shock-related protein, HSP20, in vascular smooth muscles is associated with changes in the macromolecular associations of HSP20. J Biol Chem 274, 6324-9. Brophy, C. M., Lamb, S. and Graham, A. (1999b). The small heat shock-related protein20 is an actin-associated protein. J Vasc Surg 29, 326-33. Brophy, C. M., Woodrum, D., Dickinson, M. and Beall, A. (1998). Thrombin activates MAPKAP2 kinase in vascular smooth muscle. J Vasc Surg 27, 963-9. Bukach, O. V., Marston, S. B. and Gusev, N. B. (2005). Small heat shock protein with apparent molecular mass 20 kDa (Hsp20, HspB6) is not a genuine actin-binding protein. J Muscle Res Cell Motil 26, 175-81. Bukach, O. V., Seit-Nebi, A. S., Marston, S. B. and Gusev, N. B. (2004). Some properties of human small heat shock protein Hsp20 (HspB6). Eur J Biochem 271, 291-302. Chernik, I. S., Seit-Nebi, A. S., Marston, S. B. and Gusev, N. B. (2007). Small heat shock protein Hsp20 (HspB6) as a partner of 14-3-3gamma. Mol Cell Biochem 295, 9-17. 73

Datta, S. A. and Rao, C. M. (2000). Packing-induced conformational and functional changes in the subunits of alpha -crystallin. J Biol Chem 275, 41004-10. Doppler, H., Storz, P., Li, J., Comb, M. J. and Toker, A. (2005). A phosphorylation statespecific antibody recognizes Hsp27, a novel substrate of protein kinase D. J Biol Chem 280, 15013-9. Dudich, I. V., Zav'yalov, V. P., Pfeil, W., Gaestel, M., Zav'yalova, G. A., Denesyuk, A. I. and Korpela, T. (1995). Dimer structure as a minimum cooperative subunit of small heat-shock proteins. Biochim Biophys Acta 1253, 163-8. Ehrnsperger, M., Graber, S., Gaestel, M. and Buchner, J. (1997). Binding of non-native protein to Hsp25 during heat shock creates a reservoir of folding intermediates for reactivation. Embo J 16, 221-9. Ehrnsperger, M., Lilie, H., Gaestel, M. and Buchner, J. (1999). The dynamics of Hsp25 quaternary structure. Structure and function of different oligomeric species. J Biol Chem 274, 14867-74. Farahbakhsh, Z. T., Huang, Q. L., Ding, L. L., Altenbach, C., Steinhoff, H. J., Horwitz, J. and Hubbell, W. L. (1995). Interaction of alpha-crystallin with spin-labeled peptides. Biochemistry 34, 509-16. Fontaine, J. M., Sun, X., Benndorf, R. and Welsh, M. J. (2005). Interactions of HSP22 (HSPB8) with HSP20, alphaB-crystallin, and HSPB3. Biochem Biophys Res Commun 337, 1006-11. Friedrich, K. L., Giese, K. C., Buan, N. R. and Vierling, E. (2004). Interactions between small heat shock protein subunits and substrate in small heat shock protein-substrate complexes. J Biol Chem 279, 1080-9. Fu, L. and Liang, J. J. (2003). Enhanced stability of alpha B-crystallin in the presence of small heat shock protein Hsp27. Biochem Biophys Res Commun 302, 710-4. Fuchs, L. C., Giulumian, A. D., Knoepp, L., Pipkin, W., Dickinson, M., Hayles, C. and Brophy, C. (2000). Stress causes decrease in vascular relaxation linked with altered phosphorylation of heat shock proteins. Am J Physiol Regul Integr Comp Physiol 279, R492-8. Ganea, E. (2001). Chaperone-like activity of alpha-crystallin and other small heat shock proteins. Curr Protein Pept Sci 2, 205-25. Haley, D. A., Horwitz, J. and Stewart, P. L. (1998). The small heat-shock protein, alphaB-crystallin, has a variable quaternary structure. J Mol Biol 277, 27-35. Haslbeck, M., Walke, S., Stromer, T., Ehrnsperger, M., White, H. E., Chen, S., Saibil, H. R. and Buchner, J. (1999). Hsp26: a temperature-regulated chaperone. Embo J 18, 6744-51. Horwitz, J., Huang, Q. and Ding, L. (2004). The native oligomeric organization of alphacrystallin, is it necessary for its chaperone function? Exp Eye Res 79, 817-21. Ibitayo, A. I., Sladick, J., Tuteja, S., Louis-Jacques, O., Yamada, H., Groblewski, G., Welsh, M. and Bitar, K. N. (1999). HSP27 in signal transduction and association with contractile proteins in smooth muscle cells. Am J Physiol 277, G445-54. Ito, H., Okamoto, K., Nakayama, H., Isobe, T. and Kato, K. (1997). Phosphorylation of alphaB-crystallin in response to various types of stress. J Biol Chem 272, 29934-41.

74

Kato, K., Goto, S., Inaguma, Y., Hasegawa, K., Morishita, R. and Asano, T. (1994). Purification and characterization of a 20-kDa protein that is highly homologous to alpha B crystallin. J Biol Chem 269, 15302-9. Kato, K., Ito, H., Kamei, K., Inaguma, Y., Iwamoto, I. and Saga, S. (1998). Phosphorylation of alphaB-crystallin in mitotic cells and identification of enzymatic activities responsible for phosphorylation. J Biol Chem 273, 28346-54. Kato, K., Shinohara, H., Goto, S., Inaguma, Y., Morishita, R. and Asano, T. (1992). Copurification of small heat shock protein with alpha B crystallin from human skeletal muscle. J Biol Chem 267, 7718-25. Kim, K. K., Kim, R. and Kim, S. H. (1998). Crystal structure of a small heat-shock protein. Nature 394, 595-9. Kim, M. V., Seit-Nebi, A. S., Marston, S. B. and Gusev, N. B. (2004). Some properties of human small heat shock protein Hsp22 (H11 or HspB8). Biochem Biophys Res Commun 315, 796-801. Knock, G. A., De Silva, A. S., Snetkov, V. A., Siow, R., Thomas, G. D., Shiraishi, M., Walsh, M. P., Ward, J. P. and Aaronson, P. I. (2005). Modulation of PGF2alpha- and hypoxiainduced contraction of rat intrapulmonary artery by p38 MAPK inhibition: a nitric oxidedependent mechanism. Am J Physiol Lung Cell Mol Physiol 289, L1039-48. Knoepp, L., Beall, A., Woodrum, D., Mondy, J. S., Shaver, E., Dickinson, M. and Brophy, C. M. (2000). Cellular stress inhibits vascular smooth muscle relaxation. J Vasc Surg 31, 343-53. Lambert, H., Charette, S. J., Bernier, A. F., Guimond, A. and Landry, J. (1999). HSP27 multimerization mediated by phosphorylation-sensitive intermolecular interactions at the amino terminus. J Biol Chem 274, 9378-85. Larsen, J. K., Yamboliev, I. A., Weber, L. A. and Gerthoffer, W. T. (1997). Phosphorylation of the 27-kDa heat shock protein via p38 MAP kinase and MAPKAP kinase in smooth muscle. Am J Physiol 273, L930-40. Lee, G. J., Roseman, A. M., Saibil, H. R. and Vierling, E. (1997). A small heat shock protein stably binds heat-denatured model substrates and can maintain a substrate in a foldingcompetent state. Embo J 16, 659-71. Lee, Y. J., Lee, D. H., Cho, C. K., Bae, S., Jhon, G. J., Lee, S. J., Soh, J. W. and Lee, Y. S. (2005). HSP25 inhibits protein kinase C delta-mediated cell death through direct interaction. J Biol Chem 280, 18108-19. Lelj-Garolla, B. and Mauk, A. G. (2006). Self-association and chaperone activity of Hsp27 are thermally activated. J Biol Chem 281, 8169-74. Lindner, R. A., Carver, J. A., Ehrnsperger, M., Buchner, J., Esposito, G., Behlke, J., Lutsch, G., Kotlyarov, A. and Gaestel, M. (2000). Mouse Hsp25, a small shock protein. The role of its C-terminal extension in oligomerization and chaperone action. Eur J Biochem 267, 192332. Lindner, R. A., Kapur, A. and Carver, J. A. (1997). The interaction of the molecular chaperone, alpha-crystallin, with molten globule states of bovine alpha-lactalbumin. J Biol Chem 272, 27722-9.

75

McLemore, E. C., Tessier, D. J., Thresher, J., Komalavilas, P. and Brophy, C. M. (2005). Role of the small heat shock proteins in regulating vascular smooth muscle tone. J Am Coll Surg 201, 30-6. Meier, M., King, G. L., Clermont, A., Perez, A., Hayashi, M. and Feener, E. P. (2001). Angiotensin AT(1) receptor stimulates heat shock protein 27 phosphorylation in vitro and in vivo. Hypertension 38, 1260-5. Nakajima, K., Hirade, K., Ishisaki, A., Matsuno, H., Suga, H., Kanno, Y., Shu, E., Kitajima, Y., Katagiri, Y. and Kozawa, O. (2005). Akt regulates thrombin-induced HSP27 phosphorylation in aortic smooth muscle cells: function at a point downstream from p38 MAP kinase. Life Sci 77, 96-107. Narberhaus, F. (2002). Alpha-crystallin-type heat shock proteins: socializing minichaperones in the context of a multichaperone network. Microbiol Mol Biol Rev 66, 64-93; table of contents. Panasenko, O. O., Kim, M. V., Marston, S. B. and Gusev, N. B. (2003). Interaction of the small heat shock protein with molecular mass 25 kDa (hsp25) with actin. Eur J Biochem 270, 892-901. Pivovarova, A. V., Chebotareva, N. A., Chernik, I. S., Gusev, N. B. and Levitsky, D. I. (2007). Small heat shock protein Hsp27 prevents heat-induced aggregation of F-actin by forming soluble complexes with denatured actin. Febs J 274, 5937-48. Pivovarova, A. V., Mikhailova, V. V., Chernik, I. S., Chebotareva, N. A., Levitsky, D. I. and Gusev, N. B. (2005). Effects of small heat shock proteins on the thermal denaturation and aggregation of F-actin. Biochem Biophys Res Commun 331, 1548-53. Rane, M. J., Pan, Y., Singh, S., Powell, D. W., Wu, R., Cummins, T., Chen, Q., McLeish, K. R. and Klein, J. B. (2003). Heat shock protein 27 controls apoptosis by regulating Akt activation. J Biol Chem 278, 27828-35. Rogalla, T., Ehrnsperger, M., Preville, X., Kotlyarov, A., Lutsch, G., Ducasse, C., Paul, C., Wieske, M., Arrigo, A. P., Buchner, J. and Gaestel, M. (1999). Regulation of Hsp27 oligomerization, chaperone function, and protective activity against oxidative stress/tumor necrosis factor alpha by phosphorylation. J Biol Chem 274, 18947-56. Schuck, P. (2000). Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling. Biophys J 78, 1606-19. Shi, B., Grahn, J. C., Reilly, D. A., Dizon, T. C. and Isseroff, R. R. (2008). Responses of the 27-kDa heat shock protein to UVB irradiation in human epidermal melanocytes. Exp Dermatol 17, 108-14. Sobott, F., Benesch, J. L., Vierling, E. and Robinson, C. V. (2002). Subunit exchange of multimeric protein complexes. Real-time monitoring of subunit exchange between small heat shock proteins by using electrospray mass spectrometry. J Biol Chem 277, 38921-9. Sreelakshmi, Y., Santhoshkumar, P., Bhattacharyya, J. and Sharma, K. K. (2004). AlphaA-crystallin interacting regions in the small heat shock protein, alphaB-crystallin. Biochemistry 43, 15785-95. Stamler, R., Kappe, G., Boelens, W. and Slingsby, C. (2005). Wrapping the alphacrystallin domain fold in a chaperone assembly. J Mol Biol 353, 68-79. Stromer, T., Ehrnsperger, M., Gaestel, M. and Buchner, J. (2003). Analysis of the interaction of small heat shock proteins with unfolding proteins. J Biol Chem 278, 18015-21. 76

Studer, S. and Narberhaus, F. (2000). Chaperone activity and homo- and hetero-oligomer formation of bacterial small heat shock proteins. J Biol Chem 275, 37212-8. Sugiyama, Y., Suzuki, A., Kishikawa, M., Akutsu, R., Hirose, T., Waye, M. M., Tsui, S. K., Yoshida, S. and Ohno, S. (2000). Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation. J Biol Chem 275, 1095-104. Sun, T. X., Akhtar, N. J. and Liang, J. J. (1998). Subunit exchange of lens alphacrystallin: a fluorescence energy transfer study with the fluorescent labeled alphaA-crystallin mutant W9F as a probe. FEBS Lett 430, 401-4. Sun, T. X. and Liang, J. J. (1998). Intermolecular exchange and stabilization of recombinant human alphaA- and alphaB-crystallin. J Biol Chem 273, 286-90. Sun, X., Fontaine, J. M., Rest, J. S., Shelden, E. A., Welsh, M. J. and Benndorf, R. (2004). Interaction of human HSP22 (HSPB8) with other small heat shock proteins. J Biol Chem 279, 2394-402. Sun, X., Welsh, M. J. and Benndorf, R. (2006). Conformational changes resulting from pseudophosphorylation of mammalian small heat shock proteins--a two-hybrid study. Cell Stress Chaperones 11, 61-70. Tanabe, K., Takai, S., Matsushima-Nishiwaki, R., Kato, K., Dohi, S. and Kozawa, O. (2008). alpha(2) adrenoreceptor agonist regulates protein kinase C-induced heat shock protein 27 phosphorylation in C6 glioma cells. J Neurochem. Taylor, R. P. and Benjamin, I. J. (2005). Small heat shock proteins: a new classification scheme in mammals. J Mol Cell Cardiol 38, 433-44. Theriault, J. R., Lambert, H., Chavez-Zobel, A. T., Charest, G., Lavigne, P. and Landry, J. (2004). Essential role of the NH2-terminal WD/EPF motif in the phosphorylation-activated protective function of mammalian Hsp27. J Biol Chem 279, 23463-71. van Boekel, M. A., de Lange, F., de Grip, W. J. and de Jong, W. W. (1999). Eye lens alphaA- and alphaB-crystallin: complex stability versus chaperone-like activity. Biochim Biophys Acta 1434, 114-23. van de Klundert, F. A., Smulders, R. H., Gijsen, M. L., Lindner, R. A., Jaenicke, R., Carver, J. A. and de Jong, W. W. (1998). The mammalian small heat-shock protein Hsp20 forms dimers and is a poor chaperone. Eur J Biochem 258, 1014-21. van Montfort, R. L., Basha, E., Friedrich, K. L., Slingsby, C. and Vierling, E. (2001). Crystal structure and assembly of a eukaryotic small heat shock protein. Nat Struct Biol 8, 102530. Wieske, M., Benndorf, R., Behlke, J., Dolling, R., Grelle, G., Bielka, H. and Lutsch, G. (2001). Defined sequence segments of the small heat shock proteins HSP25 and alphaBcrystallin inhibit actin polymerization. Eur J Biochem 268, 2083-90. Woodrum, D. A., Brophy, C. M., Wingard, C. J., Beall, A. and Rasmussen, H. (1999). Phosphorylation events associated with cyclic nucleotide-dependent inhibition of smooth muscle contraction. Am J Physiol 277, H931-9. Zantema, A., Verlaan-De Vries, M., Maasdam, D., Bol, S. and van der Eb, A. (1992). Heat shock protein 27 and alpha B-crystallin can form a complex, which dissociates by heat shock. J Biol Chem 267, 12936-41.

77

Zoubeidi, A., Zardan, A., Beraldi, E., Fazli, L., Sowery, R., Rennie, P., Nelson, C. and Gleave, M. (2007). Cooperative interactions between androgen receptor (AR) and heat-shock protein 27 facilitate AR transcriptional activity. Cancer Res 67, 10455-65.

78