Initial rate; Enzyme kinetics; Competitive and non-competitve models
Abstract :
[en] Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon plots can only be used when a true initial rate is measured. Despite the fact that this point has often been stressed, it is far too often ignored in favour of restricting the essay time to one where low amounts of substrate are used. When one or several irreversible and slow steps occur with an inactivator during the incubation of a ternary enzyme-substrate-inactivator mixture, the rate of the enzyme-catalyzed reaction progressively decreases. Even under these conditions, the present computer simulations investigations show that apparently linear Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon graphs can be obtained when the amount of product is mistakenly assumed to represent the true initial rate. Moreover, the observed pattern can change with time, going for instance from non-competitive to competitive. "Ki's" measured under these conditions also vary with time and bear little relationship to the true constants involved in the interaction.
Research center :
Laboratoire de Dynamique Moléculaire
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Frère, Jean-Marie ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Leyh, Bernard ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de dynamique moléculaire
Renard, André
Language :
English
Title :
Lineweaver-Burk, Hanes, Eadie-Hofstee and Dixon Plots in Non-steady-state Situations.
Publication date :
1983
Journal title :
Journal of Theoretical Biology
ISSN :
0022-5193
eISSN :
1095-8541
Publisher :
Elsevier, Amsterdam, Netherlands
Volume :
101
Pages :
387-400
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
National Institutes of Health - NIH (Contract R01 13364-05), FNRSM (Contract 3.4501.79), AC Brussels (Convention 79/81-11)
Cha, Agarwal, Parks Biochem. Pharmacol 1975, 24:2187.
Frère, Leyh-Bouille, Ghuysen, Perkins (1974) Interaction between beta-Lactam Antibiotics and Exocellular dd-Carboxypeptidase-Transpeptidase of Streptomyces R61. European Journal of Biochemistry 50:203.
Frère, Ghuysen, Reynolds, Moreno, Perkins (1974) Binding of beta-lactam antibiotics to the exocellular DD-carboxypeptidase-transpeptidase of Streptomyces R39. Biochem J 143:241.
Frére, Ghuysen, Iwatsubo (1975) Kinetics of Interaction between the Exocellular DD-Carboxypeptidase-Transpeptidase from Streptomyces R61 and beta-Lactam Antibiotics. A Choice of Models. European Journal of Biochemistry 57:343.
Frère, Ghuysen, Perkins (1975) Interaction between the Exocellular DD-Carboxypeptidase-Transpeptidase from Streptomyces R61, Substrate and beta-Lactam Antibiotics. A Choice of Models. European Journal of Biochemistry 57:353.
Frère, Ghuysen, Degelaen, Loffet, Perkins (1975) Fragmentation of benzylpenicillin after interaction with the exocellular DD-carboxypeptidase-transpeptidases of Streptomyces R61 and R39. Nature 258:168.
Frieden J. biol. Chem 1970, 245:5788.
Henderson Biochem. J 1972, 127:321.
Izaki, Strominger J. biol. Chem 1968, 243:3193.
Ketelslegers Thèse d'Agrégation, Université de Liège, Belgium; 1982.
Martin, Maskos, Burger Eur. J. Biochem 1975, 55:465.
Morrisson (1969) Kinetics of the reversible inhibition of enzyme-catalysed reactions by tight-binding inhibitors. Biochimica et Biophysica Acta (BBA) - Enzymology 185:269.