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. Study holds ProTherm entries: 3114, 3115, 14157 Extra Details: Escherichia coli adenylate kinase; catalytic role; arginine;,site-directed mutagenesis; structural

Submission Details

ID: Mohf28qL3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:21 p.m.

Version: 1

Publication Details
Reinstein J;Gilles AM;Rose T;Wittinghofer A;Saint Girons I;Bârzu O;Surewicz WK;Mantsch HH,J. Biol. Chem. (1989) Structural and catalytic role of arginine 88 in Escherichia coli adenylate kinase as evidenced by chemical modification and site-directed mutagenesis. PMID:2542263
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
6F7U 2018-03-14 1.4 Molecular Mechanism of ATP versus GTP Selectivity of Adenylate Kinase
4X8L 2015-07-22 1.7 Crystal structure of E. coli Adenylate kinase P177A mutant in complex with inhibitor Ap5a
1E4V 2000-08-04 1.85 Mutant G10V of adenylate kinase from E. coli, modified in the Gly-loop
1E4Y 2000-08-04 1.85 Mutant P9L of adenylate kinase from E. coli, modified in the Gly-loop
5EJE 2016-11-09 1.9 Crystal structure of E. coli Adenylate kinase G56C/T163C double mutant in complex with Ap5a
1ANK 1994-05-31 2.0 THE CLOSED CONFORMATION OF A HIGHLY FLEXIBLE PROTEIN: THE STRUCTURE OF E. COLI ADENYLATE KINASE WITH BOUND AMP AND AMPPNP
3HPQ 2009-11-03 2.0 Crystal structure of wild-type adenylate kinase from E. coli, in complex with Ap5A
1AKE 1994-01-31 2.0 STRUCTURE OF THE COMPLEX BETWEEN ADENYLATE KINASE FROM ESCHERICHIA COLI AND THE INHIBITOR AP5A REFINED AT 1.9 ANGSTROMS RESOLUTION: A MODEL FOR A CATALYTIC TRANSITION STATE
3HPR 2009-11-03 2.0 Crystal structure of V148G adenylate kinase from E. coli, in complex with Ap5A
4X8O 2015-07-15 2.1 Crystal structure of E. coli Adenylate kinase Y171W mutant in complex with inhibitor Ap5a
4AKE 1996-06-10 2.2 ADENYLATE KINASE
4X8H 2015-07-22 2.5 Crystal structure of E. coli Adenylate kinase P177A mutant
4X8M 2015-07-15 2.6 Crystal structure of E. coli Adenylate kinase Y171W mutant
2ECK 1997-03-12 2.8 STRUCTURE OF PHOSPHOTRANSFERASE

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
92.1 Adenylate kinase A8GAV3 KAD_SERP5
95.8 Adenylate kinase A8AJW9 KAD_CITK8
96.3 Adenylate kinase P0A1V4 KAD_SALTY
96.3 Adenylate kinase P0A1V5 KAD_SALTI
96.3 Adenylate kinase B4TMG6 KAD_SALSV
96.3 Adenylate kinase B5BD44 KAD_SALPK
96.3 Adenylate kinase C0Q812 KAD_SALPC
96.3 Adenylate kinase Q5PFK8 KAD_SALPA
96.3 Adenylate kinase B4SWY1 KAD_SALNS
96.3 Adenylate kinase B4T9I1 KAD_SALHS
96.3 Adenylate kinase B5R612 KAD_SALG2
96.3 Adenylate kinase B5QU77 KAD_SALEP
96.3 Adenylate kinase B5FLJ7 KAD_SALDC
96.3 Adenylate kinase Q57S76 KAD_SALCH
96.3 Adenylate kinase B5EXN0 KAD_SALA4
95.8 Adenylate kinase A4W7F8 KAD_ENT38
96.3 Adenylate kinase A6T5N7 KAD_KLEP7
96.3 Adenylate kinase B5Y0N3 KAD_KLEP3
99.1 Adenylate kinase B7MDZ6 KAD_ECO45
99.1 Adenylate kinase Q3Z4S5 KAD_SHISS
99.5 Adenylate kinase Q83M40 KAD_SHIFL
99.5 Adenylate kinase Q8FK84 KAD_ECOL6
99.5 Adenylate kinase Q0TKG7 KAD_ECOL5
99.5 Adenylate kinase B7MQI9 KAD_ECO81
99.5 Adenylate kinase B7NIF6 KAD_ECO7I
99.5 Adenylate kinase B7UKF4 KAD_ECO27
99.5 Adenylate kinase Q32J54 KAD_SHIDS
99.5 Adenylate kinase B7M3W6 KAD_ECO8A
100.0 Adenylate kinase Q0T7B1 KAD_SHIF8
100.0 Adenylate kinase Q325C2 KAD_SHIBS
100.0 Adenylate kinase B2U4S7 KAD_SHIB3
100.0 Adenylate kinase B7LV13 KAD_ESCF3
100.0 Adenylate kinase B1LJN2 KAD_ECOSM
100.0 Adenylate kinase B6I0C6 KAD_ECOSE
100.0 Adenylate kinase B7N927 KAD_ECOLU
100.0 Adenylate kinase P69441 KAD_ECOLI
100.0 Adenylate kinase B1IZC0 KAD_ECOLC
100.0 Adenylate kinase A7ZXD2 KAD_ECOHS
100.0 Adenylate kinase B1XFR1 KAD_ECODH
100.0 Adenylate kinase C4ZUS8 KAD_ECOBW
100.0 Adenylate kinase P69442 KAD_ECO57
100.0 Adenylate kinase B7L799 KAD_ECO55
100.0 Adenylate kinase A7ZIN4 KAD_ECO24