Le. Greene et al., EFFECT OF NUCLEOTIDE ON THE BINDING OF PEPTIDES TO 70-KDA HEAT-SHOCK PROTEIN, The Journal of biological chemistry, 270(7), 1995, pp. 2967-2973
In previous work we found that bovine brain hsp70 has a single binding
site for nucleotide, and that, with ATP at this site, the rates of as
sociation and dissociation of clathrin from hsp70 are fast, whereas wi
th ADP at this site, these rates are unmeasurably slow. In the present
study we show, first, that peptide C, cytochrome c peptide, and RNase
S peptide bind competitively with clathrin, suggesting that they bind
to the same site on hsp70, although RNase S peptide binds an order of
magnitude more weakly than peptide C and cytochrome c peptide. Second
, we show that, with ADP bound to hsp70, as occurs with clathrin, the
rate constant for dissociation of peptide markedly decreases compared
to the rate constant observed in ATP. In contrast, ADP only slightly d
ecreases the rate of association of peptide. Based on these data we pr
opose a model in which substrates of hsp70 bind to and dissociate from
the ATP form of the enzyme, while, following ATP hydrolysis, they are
locked onto the ADP form of the enzyme, unable to dissociate until AD
P is released and ATP rebinds.