Kinetic characterization of branched chain ketoacid dehydrogenase
Abstract
Initial velocity and product inhibition experiments were performed to characterize the kinetic mechanism of branched chain ketoacid dehydrogenase (the branched chain complex) activity. The results were directly compared to predicted patterns for a three-site ping-pong mechanism. Product inhibition experiments confirmed that NADH is competitive versus NAD+ and isovaleryl CoA is competitive versus CoA. Furthermore, both NADH and isovaleryl CoA were uncompetitive versus ketoisovaleric acid. These results are consistent with a ping-pong mechanism and are similar to pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase. However, inhibition patterns for isovaleryl CoA versus NAD+ and NADH versus CoA are not consistent with a ping-pong mechanism. These patterns may result from a steric interaction between the flavoprotein and transacetylase subunits of the complex. To determine the kinetic mechanism of the substrates and feedback inhibitors (NADH and isovaleryl CoA) of the branched chain complex, it was necessary to define the interaction of the inhibitors at nonsaturating fixed substrate (CoA and NAD+) concentrations. While the competitive inhibition patterns were maintained, slope replots for NADH versus NAD+ at nonsaturating CoA concentrations were parabolic. This unexpected finding resembles a linear mixed type of inhibition where the inhibition is a combination of pure competitive and noncompetitive inhibition.
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During exercise, the branched-chain amino acids (BCAA) are preferentially oxidised to other forms of amino acids through a transamination process to become transferred into keto-acids by the rate-limiting enzyme branched-chain oxo-acid dehydrogenase (BCOAD).
“During exercise, the branched-chain amino acids (BCAA) are preferentially oxidised to other forms of amino acids through a transamination process to become transferred into keto-acids by the rate-limiting enzyme branched-chain oxo-acid dehydrogenase (BCOAD) [171,172].”
From Regulation of Energy Substrate Metabolism in Endurance Exercise (2021) · cites this paper as [172]
Cites
PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT
Lowry, O. H., Rosebrough, N., Farr, A. L., & Randall, R. (1951) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)52451-6 · Cited by 315,771 · see citing works
The kinetics of enzyme-catalyzed reactions with two or more substrates or products
Cleland, W. W. (1963) · Biochimica et Biophysica Acta (BBA) - Specialized Section on Enzymological Subjects · doi:10.1016/0926-6569(63)90211-6 · Cited by 1,911 · see citing works
Enzyme Kinetics Behavior and analysis of rapid equilibrium and steady state enzyme systems
Gutfreund, H. (1975) · FEBS Letters · doi:10.1016/0014-5793(75)80457-1 · Cited by 1,508 · see citing works
α-Keto Acid Dehydrogenase Complexes
Tsai, C. S., Bürgett, M. W., & Reed, L. J. (1973) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)43138-4 · Cited by 152 · see citing works
Smith, C. M., Bryła, J., & WILLIAMSON, J. R. (1974) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)42910-4 · Cited by 128 · see citing works
Ziegler, D. M., & Pettit, F. H. (1966) · Biochemistry · doi:10.1021/bi00873a024 · Cited by 126 · see citing works
The mammalian 2-oxoacid dehydrogenases: a complex family
Yeaman, S. J. (1986) · Trends in Biochemical Sciences · doi:10.1016/0968-0004(86)90033-2 · Cited by 106 · see citing works
Partial purification and properties of branched-chain 2-oxo acid dehydrogenase of ox liver
Parker, P. J., & Randle, P. J. (1978) · Biochemical Journal · doi:10.1042/bj1710751 · Cited by 102 · see citing works
Danner, D. J., Lemmon, S. K., Besharse, J. C., et al. (1979) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)50626-8 · Cited by 97 · see citing works
Branched chain 2‐oxo‐acid dehydrogenase complex of rat liver
Parker, P. J., & Randle, P. J. (1978) · FEBS Letters · doi:10.1016/0014-5793(78)80325-1 · Cited by 75 · see citing works
Cleland, W. W. (1973) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)43139-6 · Cited by 73 · see citing works
A convenient computer program for fitting enzymatic rate laws to steady-state data
Kohn, M. C., Menten, L. E., & Garfinkel, D. J. (1979) · Computers and Biomedical Research · doi:10.1016/0010-4809(79)90032-6 · Cited by 65 · see citing works
A Kinetic Study of the α-Keto Acid Dehydrogenase Complexes from Pig Heart Mitochondria1
HAMADA, M., Koike, K., NAKAULA, Y., et al. (1975) · The Journal of Biochemistry · doi:10.1093/oxfordjournals.jbchem.a130805 · Cited by 54 · see citing works
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Tarnopolsky, M. (2004) · Nutrition · doi:10.1016/j.nut.2004.04.008 · Cited by 242 · see citing works
McKenzie, S., Phillips, S. M., Carter, S. L., Lowther, S., Gibala, M. J., & Tarnopolsky, M. A. (2000) · American Journal of Physiology-Endocrinology and Metabolism · doi:10.1152/ajpendo.2000.278.4.e580 · Cited by 219 · see citing works
Regulation of Energy Substrate Metabolism in Endurance Exercise
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- Authors
- Boyer, B., & Odessey, R.
- Affiliations
- Louisiana State University (US)
- Corresponding author
- Boyer, Bert
- ORCID
- Boyer, Bert
- Venue
- Archives of Biochemistry and Biophysics
- Publisher
- Elsevier BV
- Published
- 1991-02
- Volume
- 285
- Issue
- 1
- Pages
- 1-7
- ISSN
- 0003-9861 (ISSN-L)
- OpenAlex
- W2149201842
- Cited by
- 22
- References
- 13
- Citations by year
- 2025: 2 · 2024: 1 · 2021: 2 · 2017: 1
- FWCI
- 0.22
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Archives of Biochemistry and Biophysics (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5p01gm032654-10, 1r01gm037217-01a1, 5t32hl007098-15)
- National Heart, Lung, and Blood Institute (HL07098)
- National Institute of General Medical Sciences (GM37217, GM 32654)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Biochemical Acid Research Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Metabolism and Genetic Disorders
- Keywords
- branched-chain ketoacid dehydrogenase, enzyme kinetic mechanism, ping-pong mechanism, product inhibition, competitive inhibition, uncompetitive inhibition, NADH, flavoprotein, steric interactions, pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase
- MeSH terms
- Animals, Binding, Competitive, Flavoproteins, Ketone Oxidoreductases, Kinetics, Mitochondria, Multienzyme Complexes, NAD, Rats, Inbred Strains, 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide), Rats
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