The protein-protein complex between pp60v-src and hsp90 is stabilized by molybdate, vanadate, tungstate, and an endogenous cytosolic metal.
In a recent study demonstrating the cell-free reconstitution of the pp60v-src-hsp90 complex, we found that the tyrosine kinase-hsp90 complex is stabilized by molybdate (Hutchison, K. A., Brott, B. K., De Leon, J. H., Perdew, G. H., Jove, R., and Pratt, W. B. (1992) J. Biol. Chem. 267, 2902-2908). In this paper, we examine in detail the stabilization of this protein-protein interaction by transition metal oxyanions. The pp60v-src-hsp90 complex is stabilized by sodium molybdate with the same concentration dependence as the glucocorticoid receptor-hsp90 complex. As with the steroid receptor heterocomplexes, vanadate and tungstate also stabilize the pp60v-src-hsp90 interaction. Passage of cytosol through a Chelex-100 metal-chelating resin destabilizes the native pp60v-src-hsp90 complex, suggesting that the complex is normally stabilized by an endogenous metal factor. Readdition of either the heat-stable components of cytosol or a partially purified endogenous metal factor stabilizes the metal-depleted complex. Molybdate also stabilizes the presence of p50, a known hsp90-associated protein, in the pp60v-src heterocomplex. Given the identical effects of transition metal oxyanions on both pp60v-src- and steroid receptor-hsp90 complexes and the lack of any sequence homology between pp60v-src and the receptors, it seems very likely that it is the common component, hsp90, that contains the site of the metal interaction.
Hutchison
K A
KA
Department of Pharmacology, University of Michigan Medical School, Ann Arbor 48109.
Stancato
L F
LF
Jove
R
R
Pratt
W B
WB
D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D007739:L Cells; D046911:Macromolecular Substances; D008670:Metals / Q000494:pharmacology*; D051379:Mice; D008982:Molybdenum / Q000494:pharmacology*; D015688:Oncogene Protein pp60(v-src) / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011137:Polystyrenes / Q000494:pharmacology; D011145:Polyvinyls / Q000494:pharmacology; D011485:Protein Binding; D051381:Rats; D014162:Transfection; D014414:Tungsten / Q000494:pharmacology*; D017973:Tungsten Compounds; D014638:Vanadates / Q000494:pharmacology*
D002614:Chelating Agents; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D008670:Metals; D011137:Polystyrenes; D011145:Polyvinyls; D017973:Tungsten Compounds; C044659:molybdate; D014638:Vanadates; C006960:chelex; D008982:Molybdenum; D015688:Oncogene Protein pp60(v-src); C045951:tungstate; D014414:Tungsten