Prosomes and heat shock complexes in Drosophila melanogaster cells.
Prosomes and heat shock protein (HSP) complexes isolated from the cytoplasm of Drosophila cells in culture were biochemically and immunologically characterized. The two complexes were found to separate on sucrose gradients, allowing the analysis of their protein constituents by two-dimensional polyacrylamide gel electrophoresis and by reaction with anti-HSP sera and prosome-specific monoclonal antibodies. All of the prosomal proteins were found to be clearly distinct from the HSP; none of the prosomal proteins was synthesized de novo in heat shock. However, an antiprosome (anti-p27K) monoclonal antibody (mouse anti-duck) recognizing the Drosophila p29K prosomal protein allowed immunoprecipitation from a heat-shocked postmitochondrial supernatant of the crude HSP complex, including the low- and the high-molecular-weight components, in particular the 70 x 10(3)-molecular weight HSP. The highly purified small 16S HSP complex still contained this preexistent p29K prosomal protein, which thus also seems to be a metabolically stable constituent of the HSP complex. The significance of this structural and possibly functional relationship between prosomes and HSP, involving the highly ubiquitous and evolutionarily conserved prosomal protein p27/29K, remains to be elucidated.
de Sa
C M
CM
Institut Jacques Monod, Paris, France.
Rollet
E
E
de Sa
M F
MF
Tanguay
R M
RM
Best-Belpomme
M
M
Scherrer
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D005075:Biological Evolution; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000032:analysis; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003593:Cytoplasm / Q000032:analysis; D004331:Drosophila melanogaster / Q000032:analysis* / Q000166:cytology; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000032:analysis*; D015151:Immunoblotting; D008970:Molecular Weight; D010449:Peptide Mapping; D011233:Precipitin Tests; D012261:Ribonucleoproteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D006360:Heat-Shock Proteins; D012261:Ribonucleoproteins