A Steady-state Random-order Mechanism for the Oxidative Deamination of Norvaline by Glutamate Dehydrogenase

Overview

Journal Biochem J

Specialty Biochemistry

Date 1983 Apr 1

PMID 6870833

Citations 4

Authors

C LiMuti

J E Bell

Affiliations

Soon will be listed here.

Abstract

The kinetic mechanism of glutamate dehydrogenase with the monocarboxylic substrate norvaline was examined by using initial-rate steady-state kinetics and inhibition kinetics. To a first approximation the reaction mechanism can be described as a rapid-equilibrium random-order one. Binding synergism between the monocarboxylic substrate and coenzyme is not observed. Dissociation constants for NAD+ and 2-oxoglutarate calculated from the kinetic data assuming a rapid-equilibrium random-order model are in good agreement with independently obtained estimates. Lineweaver-Burk plots with varied norvaline concentration are not strictly linear, and it is concluded that a steady-state random-order model more accurately reflects the observed kinetics with norvaline as substrate.

Citing Articles

Multiple Forms of Glutamate Dehydrogenase in Animals: Structural Determinants and Physiological Implications.

Bunik V, Artiukhov A, Aleshin V, Mkrtchyan G Biology (Basel). 2016; 5(4).

PMID: 27983623 PMC: 5192433. DOI: 10.3390/biology5040053.

A novel mechanism of V-type zinc inhibition of glutamate dehydrogenase results from disruption of subunit interactions necessary for efficient catalysis.

Bailey J, Powell L, Sinanan L, Neal J, Li M, Smith T FEBS J. 2011; 278(17):3140-51.

PMID: 21749647 PMC: 3184404. DOI: 10.1111/j.1742-4658.2011.08240.x.

Ligand-induced changes in the conformational stability and flexibility of glutamate dehydrogenase and their role in catalysis and regulation.

Wacker S, Bradley M, Marion J, Bell E Protein Sci. 2010; 19(10):1820-9.

PMID: 20665690 PMC: 2998718. DOI: 10.1002/pro.459.

Negative co-operativity in glutamate dehydrogenase. Involvement of the 2-position in glutamate in the induction of conformational changes.

Bell E, LiMuti C, Renz C, Bell J Biochem J. 1985; 225(1):209-17.

PMID: 2858197 PMC: 1144571. DOI: 10.1042/bj2250209.

References

IWATSUBO M, Pantaloni D . [Regulation of the activity of glutamate dehydrogenase by effectors GTP and ADP: study by means of "stopped flow"]. Bull Soc Chim Biol (Paris). 1967; 49(11):1563-72. View

FAHIEN L, Strmecki M . Studies of gluconeogenic mitochondrial enzymes. 3. The conversion of alpha-ketoglutarate to glutamate by bovine liver mitochondrial glutamate dehydrogenase and glutamate-oxaloacetate transaminase. Arch Biochem Biophys. 1969; 130(1):468-77. DOI: 10.1016/0003-9861(69)90059-9. View

Engel P, Dalziel K . Kinetic studies of glutamate dehydrogenase with glutamate and norvaline as substrates. Coenzyme activation and negative homotropic interactions in allosteric enzymes. Biochem J. 1969; 115(4):621-31. PMC: 1185186. DOI: 10.1042/bj1150621. View

Engel P, Dalziel K . Kinetic studies of glutamate dehydrogenase. The reductive amination of 2-oxoglutarate. Biochem J. 1970; 118(3):409-19. PMC: 1179207. DOI: 10.1042/bj1180409. View

Fisher H, Bard J, Prough R . Transient-state intermediates involved in the hydride transfer step of the glutamate dehydrogenase reaction. Biochem Biophys Res Commun. 1970; 41(3):601-7. DOI: 10.1016/0006-291x(70)90055-0. View

Dalziel K, Egan R . The binding of oxidized coenzymes by glutamate dehydrogenase and the effects of glutarate and purine nucleotides. Biochem J. 1972; 126(4):975-84. PMC: 1178505. DOI: 10.1042/bj1260975. View

Prough R, Colen A, Fisher H . Spectrophotometric observation of a glutamate dehydrogenase-L-glutamate complex. Biochim Biophys Acta. 1972; 284(1):16-9. DOI: 10.1016/0005-2744(72)90040-x. View

Cross D, MCGREGOR L, Fisher H . The binding of -ketoglutarate in a binary complex and in a ternary complex with NADP + by L-glutamate dehydrogenase. Biochim Biophys Acta. 1972; 289(1):28-36. DOI: 10.1016/0005-2744(72)90104-0. View

Di Franco A, IWATSUBO M . Reaction mechanism of L-glutamate dehydrogenase. Characterization of optical and kinetic properties of various enzyme-reduced-coenzyme complexes. Eur J Biochem. 1972; 30(3):517-32. DOI: 10.1111/j.1432-1033.1972.tb02123.x. View

10.

Colen A, Prough R, Fisher H . The mechanism of glutamate dehydrogenase reaction. IV. Evidence for random and rapid binding of substrate and coenzyme in the burst phase. J Biol Chem. 1972; 247(24):7905-9. View

11.

Silverstein E, Sulebele G . Equilibrium kinetic study of the catalytic mechanism of bovine liver glutamate dehydrogenase. Biochemistry. 1973; 12(11):2164-72. DOI: 10.1021/bi00735a024. View

12.

Bell J, Dalziel K . A conformational transition of the oligomer of glutamate dehydrogenase induced by half-saturation with NAD + or NADP + . Biochim Biophys Acta. 1973; 309(1):237-42. DOI: 10.1016/0005-2744(73)90336-7. View

13.

Koberstein R, Sund H . Studies of glutamate dehydrogenase. The influence of ADP, GTP, and L-glutamate on the binding of the reduced coenzyme to beef-liver glutamate dehydrogenase. Eur J Biochem. 1973; 36(2):545-52. DOI: 10.1111/j.1432-1033.1973.tb02942.x. View

14.

Silverstein E . Equilibrium kinetic study of bovine liver glutamate dehydrogenase at high pH. Biochemistry. 1974; 13(18):3750-4. DOI: 10.1021/bi00715a021. View

15.

Di Franco A . Reaction mechanism of L-glutamate dehydrogenase. Transient complexes in the oxidative deamination of L-glutamate catalyzed by NAD(P)-dependent L-glutamate dehydrogenase. Eur J Biochem. 1974; 45(2):407-24. DOI: 10.1111/j.1432-1033.1974.tb03565.x. View

16.

Engel P, Chen S . A product-inhibition study of bovine liver glutamate dehydrogenase. Biochem J. 1975; 151(2):305-18. PMC: 1172361. DOI: 10.1042/bj1510305. View

17.

Rife J, Cleland W . Kinetic mechanism of glutamate dehydrogenase. Biochemistry. 1980; 19(11):2321-8. DOI: 10.1021/bi00552a007. View

18.

Alex S, Bell J . Dual nucleotide specificity of bovine glutamate dehydrogenase. The role of negative co-operativity. Biochem J. 1980; 191(2):299-304. PMC: 1162219. DOI: 10.1042/bj1910299. View

19.

Bailey J, Bell E, Bell J . Regulation of bovine glutamate dehydrogenase. The effects of pH and ADP. J Biol Chem. 1982; 257(10):5579-83. View

20.

STRUCK Jr J, SIZER I . The substrate specificity of glutamic acid dehydrogenase. Arch Biochem Biophys. 1960; 86:260-6. DOI: 10.1016/0003-9861(60)90415-x. View