PMID: 4897458Sep 1, 1969Paper

A substate-induced conformation change in the reaction of alkaline phosphatase from Escherichia coli

The Biochemical Journal
S E HalfordH Gutfeund

Abstract

1. Benzyl phosphonates were prepared and their potentialities as chromophoric reagents for the exploration of the substrate-binding site of Escherichia coli alkaline phosphatase were investigated. 4-Nitrobenzylphosphonate is a competitive inhibitor of the enzyme. 2-Hydroxy-5-nitrobenzylphosphonate changes its spectrum on binding to the enzyme. This spectral change is reversed when the phosphonate is displaced from the enzyme by substrate. 2. The kinetics of the reaction of 2-hydroxy-5-nitrophenylphosphonate were studied by the stopped-flow and the temperature-jump techniques. It was found that the combination of the phosphonate with the enzyme occurred in two successive and reversible steps: enzyme-phosphonate complex-formation followed by rearrangement of the complex. The spectral change is associated with the rearrangement. At pH8 in 1m-sodium chloride at 22 degrees the rate constant is 167sec.(-1) for the rearrangement of the initially formed binary complex and is 18sec.(-1) for the reverse process. 3. It has previously been proposed that the reactions of phosphatase with its substrates include a distinct step between enzyme-substrate combination and chemical catalysis. The rate constant involved could be predicted but not m...Continue Reading

Citations

May 11, 1971·European Journal of Biochemistry·M LazdunskiC Lazdunski
Nov 19, 2003·European Journal of Biochemistry·Stjepan Orhanović, Maja Pavela-Vrancic
Jan 24, 1973·Nature: New Biology·H N Fernley
Jan 1, 1974·Zeitschrift für allgemeine Mikrobiologie·A K Mattoo, Z M Shah
Dec 1, 1970·European Journal of Biochemistry·C LazdunskiM Lazdunski
Jul 1, 1979·Journal of Dental Research·C K FrankerT P Rezzo
Jan 1, 1975·Progress in Biophysics and Molecular Biology·A D Malcolm
Jan 1, 1975·Progress in Biophysics and Molecular Biology·H Gutfreund
Apr 16, 1998·Biochimica Et Biophysica Acta·L A BassoA R Walmsley
Mar 21, 1974·Biochimica Et Biophysica Acta·R J Maguire
Jan 1, 1982·The International Journal of Biochemistry·A Del ArcoM A Herraez
Nov 1, 1992·Archives of Biochemistry and Biophysics·S Kallia-RaftopoulosC Coutsogeorgopoulos
May 18, 1976·Biochemistry·J A Mattis, J S Fruton
Jun 30, 1992·Biochemistry·D A Theocharis, C Coutsogeorgopoulos
Sep 1, 1971·Archives of Biochemistry and Biophysics·J W Sperow, L G Butler

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