αB crystallin translocation and phosphorylation:: Signal transduction pathways and preconditioning in the isolated rat heart

被引:54
作者
Eaton, P [1 ]
Fuller, W [1 ]
Bell, JR [1 ]
Shattock, MJ [1 ]
机构
[1] St Thomas Hosp, Rayne Inst, Ctr Cardiovasc Biol & Med, Kings Coll London, London SE1 7EH, England
基金
英国惠康基金;
关键词
alpha B crystallin; translocation; ischemic preconditioning; heart;
D O I
10.1006/jmcc.2001.1418
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
In this program of studies we have characterized in detail the translocation (assessed by Triton-insolubility) and phosphorylation (using serine-45 or -59 phosphospecific antibodies) of alphaB crystallin during myocardial ischemia [both with or without ischemic preconditioning (IPC)]. Pharmacological activators and inhibitors allowed us to characterize the signaling pathways involved in alphaB crystallin phosphorylation during ischemia. Ischemic preconditioning alone caused 30% of the heart's alphaB crystallin pool to translocate, providing a significant translocation 'head-start' in protected tissue. This enhanced translocation is coupled with increased (3-fold) alphaB crystallin phosphorylation at both serine residues. The possible role of aB crystallin in the protection afforded by ischemic preconditioning is supported by the signal transduction data: which showed preconditioning-induced aB crystallin phosphorylation,can be blocked by tyrosine kinase inhibition (using genistein) and by p38 MAP kinase or PKC inhibition (using SB203580 or bisindolylmaleimide, respectively). The activation of both p38 MAP kinase and PKC are recognized requirements for the induction of preconditioning and their inhibition is known to block protection. Western immunoblotting analysis after isoelectric focusing electrophoresis, confirmed the observations made with the phosphospecific antibodies: but also showed that 27 +/- 4% of total cardiac crystallin was phosphorylated after 30 min of ischemia. alphaB crystallin exists as large polymeric aggregates in cardiac tissue under basal conditions (approximate to 1MDa as determined by gel filtration chromatography). We induced phosphorylation of alphaB crystallin during aerobic perfusion by the administration of phenylephrine. However this treatment did not alter the molecular aggregate size of alphaB crystallin. It appears that alphaB crystallin molecular aggregate size is not simply regulated by phosphorylation. alphaB crystallin may have a role to play in the myocardial protection induced by ischemic preconditioning, as both translocation and phosphorylation are both accelerated and enhanced by ischemic preconditioning. (C) 2001 Academic Press.
引用
收藏
页码:1659 / 1671
页数:13
相关论文
共 47 条
[31]  
Martin JL, 1997, CIRCULATION, V96, P4343
[32]  
Martin JL, 1998, CIRCULATION, V98, P620
[33]   Large unphosphorylated aggregates as the active form of hsp27 which controls intracellular reactive oxygen species and glutathione levels and generates a protection against TNFα in NIH-3T3-ras cells [J].
Mehlen, P ;
Hickey, E ;
Weber, LA ;
Arrigo, AP .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 241 (01) :187-192
[34]   Nuclear translocation of PKC zeta during ischemia and its inhibition by wortmannin, an inhibitor of phosphatidylinositol 3-kinase [J].
Mizukami, Y ;
Hirata, T ;
Yoshida, K .
FEBS LETTERS, 1997, 401 (2-3) :247-251
[35]   Delayed preconditioning of cultured adult rat cardiac myocytes: role of 70- and 90-kDa heat stress proteins [J].
Nayeem, MA ;
Hess, ML ;
Qian, YZ ;
Loesser, KE ;
Kukreja, RC .
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY, 1997, 273 (02) :H861-H868
[36]  
Ray PS, 2000, J AM COLL CARDIOL, V35, p401A
[37]   Transgene overexpression of αB crystallin confers simultaneous protection against cardiomyocyte apoptosis and necrosis during myocardial ischemia and reperfusion [J].
Ray, PS ;
Martin, JL ;
Swanson, EA ;
Otani, H ;
Dillmann, WH ;
Das, DK .
FASEB JOURNAL, 2001, 15 (02) :393-402
[38]   Regulation of Hsp27 oligomerization, chaperone function, and protective activity against oxidative stress tumor necrosis factor α by phosphorylation [J].
Rogalla, T ;
Ehrnsperger, M ;
Preville, X ;
Kotlyarov, A ;
Lutsch, G ;
Ducasse, C ;
Paul, C ;
Wieske, M ;
Arrigo, AP ;
Buchner, J ;
Gaestel, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (27) :18947-18956
[39]   Dynamic O-GlcNAcylation of the small heat shock protein alpha B-crystallin [J].
Roquemore, EP ;
Chevrier, MR ;
Cotter, RJ ;
Hart, GW .
BIOCHEMISTRY, 1996, 35 (11) :3578-3586
[40]  
Shattock MJ, 1997, CIRCULATION, V96, P311