Influence of phosphorylation and oligomerization on the protective role of the small heat shock protein 27 in rat adult cardiomyocytes.
Recent reports have demonstrated that the heat shock proteins (hsp) and in particular the hsp70 confer protection against cardiac ischemic damage. More recently, we have shown that increased expression of another heat shock protein, the hsp27, through an adenovirus vector system protects adult cardiomyocytes against ischemic injury. This small heat shock protein undergoes phosphorylation when the cell is under stress. This has led many to speculate that phosphorylation of hsp27 is required for the protective role this protein plays in the cell. In order to investigate this possibility, we have mutated the serines that are the sites of phosphorylation on the hsp27, to glycines or alanines. These nonphosphorylatable mutants of hsp27 were cloned into adenoviral vectors and used to infect adult rat cardiomyocytes to assess their ability in protecting against ischemic injury. In addition, we used a specific inhibitor of p38 MAP kinase that is a key member of the kinase pathway responsible for phosphorylating the hsp27. Our present results show that the nonphosphorylated hsp27 forms larger oligomeric complexes than the phosphorylated hsp27. Interestingly, phosphorylation of hsp27 seems not to play a role in its ability to protect adult rat cardiomyocytes against ischemic damage.
Martin
J L
JL
Department of Physiology, The Cardiovascular Institute, Loyola University, Chicago, IL, USA.
Hickey
E
E
Weber
L A
LA
Dillmann
W H
WH
Mestril
R
R
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002478:Cells, Cultured; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D020928:Mitogen-Activated Protein Kinases; D017202:Myocardial Ischemia / Q000517:prevention & control*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D048051:p38 Mitogen-Activated Protein Kinases
D006360:Heat-Shock Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases