Temperature-dependent oligomerization of hsp85 in vitro.
The failure of conventional subcellular fractionation methods to identify interactions between the bulk of hsp85 and other cellular structures suggested that critical stress protein interactions might be detectable only at elevated temperatures. This was confirmed by showing that incorporation of hsp85 and grp95 into sedimentable complexes in Triton X-100 extracts of L929 cells increased progressively over the 30 degrees C-43 degrees C temperature range. Whereas several other proteins, including hsp110 and hsp69, became sedimentable under these conditions, this effect required temperatures of approximately 43 degrees C and was only partially detergent-dependent. In contrast, hsp85 became sedimentable at temperatures as low as 33 degrees C, and this effect was highly detergent-dependent. Temperature-dependent conversion of purified hsp85 to a sedimentable form was shown to result from limited oligomerization of the protein, which occurred in the presence of detergent. Since the detergent requirement could be met by a variety of compounds, including sphingosine, these findings suggest that hsp85 oligomerization may occur when intact cells are exposed to elevated temperature.
Department of Pathology, SUNY Health Science Center, Brooklyn 11203.
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D000818:Animals; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006358:Hot Temperature; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding
D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; C018551:glucose-regulated proteins