[The balance between Hsp70 and its cochaperones Hdj1 and Bag1 determines its substrate-binding activity].
Heat shock protein Hsp70 presents one of the most effective cell protective systems. Its protective activity is mostly due to the fact that Hsp70 is able to restore native conformation of newly synthesized or damaged proteins. Two other proteins. Hdj and Bag 1, are involved in the process, allowing Hsp70 to perform binding-release cyclec of target proteins. The aim of this study was to investigate interactions between cochaperones Hdj 1 and Bag 1, and the major cell chaperone Hsp in vitro. The accumulation of Hsp70 and Hdj 1 in human erythroleukemia K562 cells was stimulated by heat stress (43 degrees C, 60 min). Cells were collected at certain time periods after heat stress, and amounts of cell chaperones were measured using Western blotting and ELISA assay. The level of Hsp70 chaperone activity in cell extracts was estimated using original technique. The effects of exogenous cochaperones and of their parts on this activity were also investigated. The results of the study indicate that Hsp70 chaperone activity is regulated by the level of its cochaperones, especially Hdj 1. At the same time the amount of ATP appears to be critical for functional activity of Hsp70. Hdj 1 and Bag 1 peptides, which bind to Hsp70 with high affinity, are able to significally reduce its chaperone activity. This finding confirms the possibility of using peptide approach for regulation of Hsp70 function at the cellular and organismal levels.
Novoselov
S S
SS
Novoselova
T V
TV
Verbova
M V
MV
Margulis
B A
BA
Guzhova
I V
IV
D000255:Adenosine Triphosphate / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D003470:Culture Media; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D020014:K562 Cells; D011485:Protein Binding; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism*
C091732:BCL2-associated athanogene 1 protein; D003470:Culture Media; D004268:DNA-Binding Proteins; C494647:DNAJB1 protein, human; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D014157:Transcription Factors; D000255:Adenosine Triphosphate