Adenylosuccinate synthase from Saccharomyces cerevisiae: homologous overexpression, purification and characterization of the recombinant protein.
Adenylosuccinate synthase (EC 6.3.4.4) catalyses the first committed step in the synthesis of adenosine. We have overexpressed the cloned gene of Saccharomyces cerevisiae (ADE12) in S. cerevisiae. The recombinant enzyme exhibits similar kinetic behaviour to that of the native enzyme purified from S. cerevisiae. This ter-reactant dimeric enzyme shows Michaelis-Menten kinetics only with IMP. l-Aspartate and GTP display a weak negative co-operativity (Hill coefficient 0. 8-0.9). This negative co-operativity has not yet been reported for adenylosuccinate synthases from other organisms. Another unusual feature of the enzyme from S. cerevisiae is its negligible inhibition by adenine nucleotides and its pronounced inhibition by Cl(-) ions.
University Bayreuth, Biochemistry II, Universitätsstrasse 30, D-95447 Bayreuth, Germany. georg.lipps@uni-bayreuth.de
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D000227:Adenine Nucleotides / Q000494:pharmacology; D000265:Adenylosuccinate Synthase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006160:Guanosine Triphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007291:Inosine Monophosphate / Q000378:metabolism; D007700:Kinetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013379:Substrate Specificity
D000227:Adenine Nucleotides; D011994:Recombinant Proteins; D007291:Inosine Monophosphate; D001224:Aspartic Acid; D006160:Guanosine Triphosphate; D000265:Adenylosuccinate Synthase