Determination of the kinetic properties of enzymes catalysing coupled reaction sequences

Duggleby R.G. (1983) Determination of the kinetic properties of enzymes catalysing coupled reaction sequences. Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular, 744 3: 249-259. doi:10.1016/0167-4838(83)90197-8


Author Duggleby R.G.
Title Determination of the kinetic properties of enzymes catalysing coupled reaction sequences
Journal name Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
ISSN 0167-4838
Publication date 1983-05-18
Sub-type Article (original research)
DOI 10.1016/0167-4838(83)90197-8
Volume 744
Issue 3
Start page 249
End page 259
Total pages 11
Subject 1303 Specialist Studies in Education
1304 Biophysics
1312 Molecular Biology
1315 Structural Biology
2700 Medicine
Abstract In a coupled system of two enzyme-catalysed reactions the concentration of the final product follows a sigmoidal process curve. The shape of this curve is dependent on the concentrations and kinetic properties of both enzymes in the system; provided that the activities of the two enzymes are similar, the kinetic properties of both enzymes may be determined simultaneously by analysis of such curves. The value of this approach is substantiated using a model system consisting of pyruvate kinase (EC 2.7.1.40) and lactate dehydrogenase (EC 1.1.1.27). Analysis of the data by a combination of numerical integration and nonlinear regression yielded values for the kinetic parameters which are in excellent agreement with those determined from measurements of steady-state rates. Appropriate experimental designs for this type of progress curve analysis are considered. The analysis is compared with, and is shown to be of much greater generality than, published procedures involving measurements of the early stages of progress curves.
Keyword Coupled enzyme reaction
Lactate dehydrogenase
Product progress curve
Pyruvate kinase
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import
 
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