DNA-directed in vitro synthesis and assembly of the form II D-ribulose-1,5-bisphosphate carboxylase/oxygenase from Rhodopseudomonas sphaeroides

Abstract

A biochemical analysis of the in vitro assembly of the form II ribulose-1,5-bisphosphate carboxylase/oxygenase from Rhodopseudomonas sphaeroides after transcription and translation from cloned DNA is presented. The predominant enzymatically active oligomeric forms of the in vitro-synthesized and -assembled ribulose-1,5-bisphosphate carboxylase are tetramers and hexamers. Assembly of the monomeric subunits to form active enzyme appears to be dependent on the presence of a minimum number of subunits in the cell extract. Assembly of ribulose-1,5-bisphosphate carboxylase also was observed when the protein-synthesizing extracts were prepared from cells which were partially derepressed for ribulose-1,5-bisphosphate carboxylase expression.