A project has been initiated to study the structure of two small heat shock proteins The sHSP's are ubiquitous among eukaryotes and are hypothesized to be a new class of molecular chaperone. The initial studies are with pea HSP 18. 1 and its S. cerevisiae homologue HSP26. Both proteins are small (18.1 kDa and -23 kDa respectively) but appear to exist as large homo-oligomers. From sedimentation and size-exclusion chromatography, the oligomer of the HSPI 8.1 could be a dodecamer and the HSP26 could be a 24-mer. Preliminary experiments are promising, although there is a large background of denatured protein film winch is common for tins type of protein. Further work will involve fixation of the protein and/or pretreatment of the carbon film substrate. We have been concentrating on the Hsp 26 winch behaves somewhat better thari the Hsp 18. 1. A brief glutaraldehyde fixation prior to deposition on the grid seems to help, but the optimal conditions have not yet been attained.
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