Molecular characterization of Escherichia coli malate synthase G. Differentiation with the malate synthase A isoenzyme.
Two genes encoding the enzymes malate synthase G and glycolate oxidase, have been linked to locus glc (64.5 min), responsible for glycolate utilization in Escherichia coli. The gene encoding malate synthase G, for which we propose the notation glcB, has been cloned, sequenced and found to correspond to a 2262-nucleotide open-reading frame, which can encode a 723-amino-acid polypeptide, clearly different from the isoenzyme malate synthase A, which has 533 amino acids. Northern-blot experiments indicate that glcB was expressed as an apparently monocistronic transcript, inducible by glycolate. Malate synthase G was purified to near homogeneity. The molecular mass determined by gel filtration yielded a value of 82 kDa for the purified enzyme and the same value as for the crude extract enzyme, indicating a monomeric structure. Despite the lower sequence similarity between malate synthase G and the other reported malate synthases, three out of nine consensus boxes defined in most of these enzymes are conserved in addition to a cysteine residue that has been reported to be important for the catalytic mechanisms.
Molina
I
I
Department of Biochemistry, School of Pharmacy, University of Barcelona, Spain.
Pellicer
M T
MT
Badia
J
J
Aguilar
J
J
Baldoma
L
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000201:enzymology; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D007700:Kinetics; D046911:Macromolecular Substances; D008292:Malate Synthase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D007527:Isoenzymes; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D008292:Malate Synthase