Enzymatic properties of chorismate synthase isozymes of tomato (Lycopersicon esculentum Mill.).
Three plastidic chorismate synthase isozymes (CS1, CS2 and CS2 delta) of tomato were identified by isolation of the corresponding cDNAs. These three cDNAs are derived from only two genes (LeCS1 and LeCS2). This additional complexity results from differential splicing of the primary transcript of one of the genes (LeCS2) giving rise to two different transcripts (CS2 and CS2 delta transcripts). All three isozymes were individually expressed in Escherichia coli both as precursor proteins with N-terminal transit peptides and as mature proteins. Only the mature but not the precursor isozymes CS1 and CS2 were enzymatically active. The enzyme CS2 delta was unstable in E. coli. Both CS1 and CS2 were purified to near homogeneity and their enzymatic properties were analyzed. They differ substantially in their Km values for the substrate 5-enol-pyruvylshikimate 3-phosphate (11 and 80 microM for the mature forms of CS1 and CS2, respectively). The two isozymes appear to be active only as oligomers, and the potential physiological implications of this result are discussed.
Braun
M
M
Institute of Plant Sciences, Swiss Federal Institute of Technology, Zürich, Switzerland.
Henstrand
J M
JM
Görlach
J
J
Amrhein
N
N
Schmid
J
J
D000595:Amino Acid Sequence; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008190:Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018551:Solanum lycopersicum / Q000201:enzymology*; D008969:Molecular Sequence Data; D019761:Phosphorus-Oxygen Lyases; D017386:Sequence Homology, Amino Acid
D007527:Isoenzymes; D008190:Lyases; C019895:chorismate synthase; D019761:Phosphorus-Oxygen Lyases