UNIQUE STRUCTURAL FEATURES OF A NOVEL CLASS OF SMALL HEAT-SHOCK PROTEINS

Small heat shock proteins (smHSPs) and alpha-crystallins constitute a family of related molecular chaperones that exhibit striking variabili ty in size, ranging from 16 to 43 kDa. Structural studies on these pro teins have been hampered by their tendency to form large, often dynami c and heterogeneous oligomeric complexes. Here we describe the structu re and expression of HSP12.6, a member of a novel class of smHSPs from the nematode Caenorhabditis elegans. Like other members of its class, HSP12.6 possesses a conserved Lu-crystallin domain but has the shorte st N- and C-terminal regions of any known smHSP. Expression of HSP12.6 is limited to the first larval stage of C. elegans and is not signifi cantly up regulated by a wide range of stressors. Unlike other smHSPs, HSP12.6 does not form large oligomeric complexes in vivo. HSP12.6 was produced in Escherichia coil as a soluble protein and purified. Cross -linking and sedimentation velocity analyses indicate that the recombi nant HSP12.6 is monomeric, making it an ideal candidate for structure determination. Interestingly, HSP12.6 does not function as a molecular chaperone in vitro, since it is unable to prevent the thermally induc ed aggregation of a test substrate. The structural and functional impl ications of these findings are discussed.