Expression and native structure of cytosolic class II small heat-shock proteins.
Higher plants synthesize small heat-shock proteins (smHSPs) from five related gene families. The class I and II families encode cytosolic smHSPs. We characterized the class II smHSPs of pea (Pisum sativum) and compared them with class I smHSPs. Antibodies against recombinant HSP17.7, a class II smHSP, recognized four heat-inducible 17- to 18-kD polypeptides and did not cross-react with class I smHSPs. On sucrose gradients the class II smHSPs sedimented primarily at 8 Svedberg units, indicating that they are components of large complexes similar in size to class I smHSP complexes. However, the class I and II complexes were readily distinguishable by nondenaturing polyacrylamide gel electrophoresis and isoelectric focusing. Nondenaturing immune precipitations using anti-HSP17.7 or anti-HSP18.1 (a class I smHSP) antiserum provide further evidence that the class I and II smHSPs exist in different complexes, composed primarily of smHSPs. Recombinant HSP17.7 and HSP18.1 formed complexes of sizes similar to those formed in vivo. When these two smHSPs were mixed, denatured with urea, and then dialyzed, the distinct class I and II complexes again formed, each containing only HSP18.1 or HSP17.7. Thus, cytosolic smHSPs from two related gene families expressed simultaneously form distinct complexes in vivo, suggesting that they have subtly different functions.
Helm
K W
KW
Department of Biology, Siena College, Loudonville, New York 12211, USA. helm@siena.edu
Lee
G J
GJ
Vierling
E
E
D000918:Antibody Specificity; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006358:Hot Temperature; D007106:Immune Sera; D018532:Pisum sativum / Q000235:genetics*; D011487:Protein Conformation
D006360:Heat-Shock Proteins; D007106:Immune Sera