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Product identification and kinetic studies of nucleotidyl cyclase activity in isolated chloroplasts by quantitative fast-atom bombardment mass spectrometry

✍ Scribed by Russell P. Newton; Mark A. Bayliss; James A. Langridge; Adam C. R. Wilkins; David E. Games; Terence J. Walton; A. Gareth Brenton; Penny E. Diffley; Frank M. Harris; Christopher J. Smith

Publisher: John Wiley and Sons
Year: 1999
Tongue: English
Weight: 106 KB
Volume: 13
Category: Article
ISSN: 0951-4198
DOI: 10.1002/(sici)1097-0231(19990615)13:11<979::aid-rcm594>3.0.co;2-p

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✦ Synopsis

While the natural occurrence of adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate in higher plants is now well established, the existence in plants of specific nucleotidyl cyclase activities functioning to synthesize these two cyclic nucleotides has been the topic of recent controversy. Here positive-ion fast-atom bombardment mass spectrometry is used to unequivocally identify the products of the putative nucleotidyl cyclase activities in a membrane fraction from isolated spinach chloroplasts, and quantitative mass spectrometry is used in a study of enzymic reaction kinetics to determine the relevant kinetic parameters necessary for the elucidation of the binding interactions between the substrates and the two enzymes.

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