Analysis of the reaction mechanism of the non‐specific endonuclease of Serratia marcescens using an artificial minimal substrate
Authors:
- Ingo FrankeIFIngo Franke
- Peter FriedhoffPFPeter Friedhoff
- Alfred PingoudAPAlfred Pingoud
- 18 November 1996
- journal article
- Published by Wiley in FEBS Letters
- Vol. 397 (2-3) , 343-346
Abstract
We have studied the mechanism of action of the Serratia nuclease using deoxythymidine 3′,5′‐bis‐(p‐nitrophenyl‐phosphate) as a substrate. A comparison of the activity with which the wild‐type enzyme and several mutant enzymes attack this artificial substrate and herring sperm DNA, respectively, supports the suggestion that His89 is the general base and a Mg2+ ion bound to Glu127 the general acid in the mechanism of phosphodiester bond hydrolysis by the Serratia nuclease, and that Asn119 directly participates in catalysis, for example by transition state stabilisation. Arg57, Arg87 and Arg131, essential for nuclease activity, are not needed for cleavage of the artificial substrate, suggesting that they are involved in binding and positioning of nucleic acid substrates.Keywords
This publication has 17 references indexed in Scilit:
- Identification of the Serratia endonuclease dimer: Structural basis and implications for catalysisProtein Science, 1996
- Sequence preferences in cleavage of dsDNA and ssDNA by the extracellular Serratia marcescens endonucleaseBiochemistry, 1995
- The Roles of Arginine 41 and Tyrosine 76 in the coupling of DNA Recognition to Phosphodiester Bond Cleavage by DNase I: a Study using Site-directed MutagenesisJournal of Molecular Biology, 1995
- 2.1 Å structure of Serratia endonuclease suggests a mechanism for binding to double-stranded DNANature Structural & Molecular Biology, 1994
- Value of General Acid-Base Catalysis to Ribonuclease AJournal of the American Chemical Society, 1994
- Nuclease structure and catalytic functionCurrent Opinion in Structural Biology, 1992
- Structure and catalytic function of nucleases: Current opinion in structural biology 1991, 1:130–138Current Opinion in Structural Biology, 1991
- The extracellular nuclease gene of Serratia marcescens and its secretion from Escherichia coliGene, 1987
- Action of staphylococcal nuclease on synthetic substratesBiochemistry, 1969
- Hammett Substituent Constants for Electron-withdrawing Substituents: Dissociation of Phenols, Anilinium Ions and Dimethylanilinium IonsJournal of the American Chemical Society, 1959