Assembly of Rubisco from native subunits.
Large subunits of ribulosebisphosphate carboxylase/oxygenase (Rubisco) (3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39) from prokaryotic sources can assemble into intact enzyme either in vitro or in Escherichia coli cells. Large subunits of higher plant Rubisco do not assemble into Rubisco in E. coli cells, nor is it possible to reconstitute higher plant Rubisco from its dissociated subunits in vitro. This behavior represents an obstacle to any practical attempts at engineering the higher plant enzyme, and it suggests that the in vivo assembly mechanism of higher plant Rubisco must be more complex than is commonly expected for oligomeric proteins of organelles. In pea chloroplasts, a binding protein interacts with newly synthesized large subunits, in quantities expected for an intermediate in the assembly process, as judged by Western blotting. Radiotracer-labeled large subunits which interact with this binding protein can be shown to assemble into Rubisco in reactions which lead to changes in the aggregation state of the binding protein. Antibody to this binding protein specifically inhibits the assembly of these subunits into Rubisco. Rubisco synthesis appears to be subject to many types of control: gene dosage, transcription rate, selective translation of message, post-translational degradation and threshold concentration effects have been observed in various organisms' synthesis of Rubisco. The biochemical mechanisms underlying most of these effects have not been elucidated. The post-translational assembly mechanism in particular appears to require further study.
Roy
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H
Biology Department, Rensselaer Polytechnic Institute, Troy, NY 12180-3590.
Cannon
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S
Gilson
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M
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology; D056890:Eukaryota / Q000201:enzymology; D005786:Gene Expression Regulation; D046911:Macromolecular Substances; D010944:Plants / Q000201:enzymology; D011499:Protein Processing, Post-Translational; D012273:Ribulose-Bisphosphate Carboxylase / Q000096:biosynthesis* / Q000235:genetics
D002352:Carrier Proteins; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D012273:Ribulose-Bisphosphate Carboxylase