title: A critical arginine in the large subunit of ribulose bisphosphate carboxylase/oxygenase identified by site-directed mutagenesis.
abstract: Rapid inactivation by phenylglyoxal of ribulose bisphosphate carboxylase/oxygenase (ribulose-P2 carboxylase) from the cyanobacterium Anacystis nidulans suggests the presence of an essential arginine, the modification of which is reduced in the presence of the substrate ribulose bisphosphate. Arginine 292 in the large subunit of ribulose-P2 carboxylase from A. nidulans was chosen for site-directed mutagenesis studies on the basis of the complete conservation of this residue in corresponding sequences of ribulose-P2 carboxylase from divergent organisms. Arginine 292 was changed to leucine and to lysine by directed mutagenesis using suitable plasmids and the bacteriophage M13. Both substitutions resulted in the production of purifiable holoenzyme with no activity after expression in Escherichia coli.
authors.affiliation: Biochemistry/Biophysics Program, Washington State University, Pullman 99164-4660.
authors.forename: R L
authors.initials: RL
authors.lastname: Haining
authors.forename: B A
authors.initials: BA
authors.lastname: McFadden
mesh_terms: D000421:Alcaligenes / Q000201:enzymology; D000595:Amino Acid Sequence; D001120:Arginine; D015153:Blotting, Western; D000460:Chlorophyta / Q000201:enzymology; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010658:Phenylglyoxal / Q000494:pharmacology; D010947:Plants, Toxic; D010957:Plasmids; D012273:Ribulose-Bisphosphate Carboxylase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014026:Nicotiana / Q000201:enzymology; D014162:Transfection; D003313:Zea mays / Q000201:enzymology
chemical_list: D001120:Arginine; D012273:Ribulose-Bisphosphate Carboxylase; D010658:Phenylglyoxal