A test for measuring the effects of enzyme inactivation.
In the single-enzyme, single-substrate reaction with non-mechanism-based enzyme inactivation, the formation of the product and inactivation of the enzyme occur independently. For this reaction, we show that the steady-state hypothesis is applicable even when degradation of the enzyme occurs. An equation for the rate of product formation has been derived and it shows Michaelis-Menten kinetics with an apparent Michaelis-Menten constant K(M)(app)=K(M)+K(delta) where K(delta) is the enzyme inactivation constant. Use of a Lineweaver-Burk plot yields values for K(M)(app), which can be used to estimate K(delta) and, consequently, the degree of enzyme inactivation in a particular experiment. We employ this methodology to estimate the inactivation constant for the arsenate reductase catalyzed production of arsenite with appreciable enzyme inactivation.
Schnell
Santiago
S
Complex Systems Group, Indiana University School of Informatics, 1900 East Tenth Street, Bloomington, IN 47406, USA. schnell@indiana.edu
Hanson
Sonya M
SM
D053502:Arsenate Reductases / Q000378:metabolism; D018053:Arsenites / Q000378:metabolism; D002384:Catalysis; D004798:Enzymes / Q000378:metabolism*; D007700:Kinetics; D008722:Methods; D011489:Protein Denaturation
D018053:Arsenites; D004798:Enzymes; D053502:Arsenate Reductases; C015001:arsenite