title: Structural and catalytic role of arginine 88 in Escherichia coli adenylate kinase as evidenced by chemical modification and site-directed mutagenesis.
abstract: Phenylglyoxal 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.
authors.affiliation: Max-Planck Institut für Medizinische Forschung, Abteilung Biophysik, Heidelberg, West Germany.
authors.forename: J
authors.initials: J
authors.lastname: Reinstein
authors.forename: A M
authors.initials: AM
authors.lastname: Gilles
authors.forename: T
authors.initials: T
authors.lastname: Rose
authors.forename: A
authors.initials: A
authors.lastname: Wittinghofer
authors.forename: I
authors.initials: I
authors.lastname: Saint Girons
authors.forename: O
authors.initials: O
authors.lastname: Bârzu
authors.forename: W K
authors.initials: WK
authors.lastname: Surewicz
authors.forename: H H
authors.initials: HH
authors.lastname: Mantsch
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000249:Adenosine Monophosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000263:Adenylate Kinase / Q000235:genetics / Q000378:metabolism*; D001120:Arginine; D002152:Calorimetry, Differential Scanning; D002384:Catalysis; D055598:Chemical Phenomena; D002621:Chemistry; D015226:Dinucleoside Phosphates / Q000494:pharmacology; D004355:Drug Stability; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D005583:Fourier Analysis; D006358:Hot Temperature; D007700:Kinetics; D003012:Microbial Collagenase / Q000378:metabolism; D009154:Mutation; D010658:Phenylglyoxal / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism*; D013055:Spectrophotometry, Infrared; D013329:Structure-Activity Relationship; D013816:Thermodynamics
chemical_list: D015226:Dinucleoside Phosphates; D000249:Adenosine Monophosphate; C012275:P(1),P(5)-di(adenosine-5'-)pentaphosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D001120:Arginine; D010770:Phosphotransferases; D000263:Adenylate Kinase; D003012:Microbial Collagenase; D010658:Phenylglyoxal