Structural and catalytic role of arginine 88 in Escherichia coli adenylate kinase as evidenced by chemical modification and site-directed mutagenesis

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TypeArticle
Journal titleThe Journal of Biological Chemistry
ISSN1083-351X
Volume264
Issue14
Pages81078112
AbstractPhenylglyoxal inactivates Escherichia coli adenylate kinase by modifying a single arginine residue (Arg-88). ATP, ADP, P1,P5-di(adenosine 5')-pentaphosphate, and to a lesser extent AMP protect the enzyme against inactivation by phenylglyoxal. Site-directed mutagenesis of Arg-88 to glycine yields a modified form of adenylate kinase (RG88 mutant) closely related structurally to the wild-type protein as indicated by Fourier transform infrared spectroscopy, differential scanning calorimetry, and limited proteolysis. However, this modified protein has only 1% of the maximum catalytic activity of the wild-type enzyme and 5- and 85-fold higher apparent Km values for ATP and AMP, respectively, than the parent adenylate kinase. Arg-88, which is a highly conserved residue in all known molecular forms of adenylate kinases (corresponding to Arg-97 in muscle cytosolic enzyme), should be located inside a big cleft of the molecule, close to the phosphate-binding loop. It possibly stabilizes the transferable gamma-phosphate group from ATP to AMP in the transition state.
Publication date
PublisherThe American Society for Biochemistry and Molecular Biology, Inc.
Linkhttp://www.jbc.org/content/264/14/8107.abstract
LanguageEnglish
AffiliationNational Research Council Canada
Peer reviewedYes
NPARC number23001352
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Record identifieracfffc56-14d0-4bf1-b320-43ea4f77f0bf
Record created2017-01-23
Record modified2017-01-23
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