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

Study Summary

Number of data points 9
Proteins Adenylate kinase ; Adenylate kinase
Unique complexes 2
Assays/Quantities/Protocols Experimental Assay: ddG ; Experimental Assay: dHcal ; Experimental Assay: Tm ; Experimental Assay: dHvH ; Derived Quantity: dTm
Libraries Mutations for sequence MRIILLGAPGAGKGTQAQFIMEKYGIPQISTGDMLRAAVKSGSELGKQAKDIMDAGKLVTDELVIALVKERIAQEDCRNGFLLDGFPRTIPQADAMKEAGINVDYVLEFDVPDELIVDRIVGRRVHAPSGRVYHVKFNPPKVEGKDDVTGEELTTRKDDQEETVRKRLVEYHQMTAPLIGYYSKEAEAGNTKYAKVDGTKPVAEVRADLEKILG

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 UniProtKB Entries

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