The small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and its precursor expressed in Escherichia coli are associated with groEL protein.
The small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase is synthesized in the cytoplasm as a precursor which is transported into the chloroplast. During or after transport the precursor is processed to its mature size by removal of an amino-terminal transit peptide. Eight small subunits and eight large subunits (synthesized in the chloroplast) assemble to form the holoenzyme. We have expressed the precursor of the small subunit in Escherichia coli as a fusion to the carboxyl terminus of staphylococcal protein A'. The fusion protein was recovered from the bacterial lysate by chromatography on IgG-agarose. A 58-kDa protein copurified with the fusion protein in approximately equal amounts. Much less of the 58-kDa protein copurified with a fusion in which the transit peptide was deleted, and it did not copurify with protein A'. The 58-kDa protein was identified as the E. coli groEL gene product with antibodies directed against a homologous mitochondrial heat shock protein. This finding is particularly interesting because a chloroplast protein involved in the assembly of ribulose-1,5-bisphosphate carboxylase/oxygenase also is homologous to the groEL protein. These homologs could modulate protein-protein interactions during folding and assembly of subunits into native complexes.
Landry
S J
SJ
Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
Bartlett
S G
SG
D001426:Bacterial Proteins / Q000302:isolation & purification; D015153:Blotting, Western; D018834:Chaperonin 60; D003001:Cloning, Molecular; D004792:Enzyme Precursors / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D006360:Heat-Shock Proteins / Q000302:isolation & purification; D046911:Macromolecular Substances; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics* / Q000302:isolation & purification
D001426:Bacterial Proteins; D018834:Chaperonin 60; D004792:Enzyme Precursors; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D012273:Ribulose-Bisphosphate Carboxylase