Ribulose 1,5-bisphosphate Carboxylase. Effect on the Catalytic Properties of Changing Methionine-330 to Leucine in the Rhodospirillum Rubrum Enzyme

Overview

Journal Biochem J

Specialty Biochemistry

Date 1986 May 1

PMID 3092806

Citations 12

Authors

B E Terzaghi

W A Laing

J T Christeller

G B Petersen

D F Hill

Affiliations

Soon will be listed here.

Abstract

Oligonucleotide-directed mutagenesis of cloned Rhodospirillum rubrum ribulose bisphosphate carboxylase/oxygenase with a synthetic 13mer oligonucleotide primer was used to effect a change at Met-330 to Leu-330. The resultant enzyme was kinetically examined in some detail and the following changes were found. The Km(CO2) increased from 0.16 to 2.35 mM, the Km(ribulose bisphosphate) increased from 0.05 to 1.40 mM for the carboxylase reaction and by a similar amount for the oxygenase reaction. The Ki(O2) increased from 0.17 to 6.00 mM, but the ratio of carboxylase activity to oxygenase activity was scarcely affected by the change in amino acid. The binding of the transition state analogue 2-carboxyribitol 1,5-bisphosphate was reversible in the mutant and essentially irreversible in the wild type enzyme. Inhibition by fructose bisphosphate, competitive with ribulose bisphosphate, was slightly increased in the mutant enzyme. These data suggest that the change of the residue from methionine to leucine decreases the stability of the enediol reaction intermediate.

Citing Articles

Structure-function studies with the unique hexameric form II ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Rhodopseudomonas palustris.

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Binding of a transition state analog to newly synthesized Rubisco.

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Missense Mutation in the Chlamydomonas Chloroplast Gene that Encodes the Rubisco Large Subunit.

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Three-dimensional structure of ribulose-1,5-bisphosphate carboxylase/oxygenase from Rhodospirillum rubrum at 2.9 A resolution.

Schneider G, Lindqvist Y, Branden C, Lorimer G EMBO J. 1986; 5(13):3409-15.

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