PMID: 9654091Jul 8, 1998

Effect of single mutations on the structural dynamics of a DNA repair enzyme, the Escherichia coli formamidopyrimidine-DNA glycosylase--a fluorescence study using tryptophan residues as reporter groups

European Journal of Biochemistry
S V KuznetsovR Santus

Abstract

The effects on the structure dynamics of the Escherichia coli wild-type formamidopyrimidine-DNA glycosylase (Fpg) protein of the single mutations Lys57-->Gly (FpgK57G), Pro2-->Gly (FpgP2G) and Pro2-->Glu (FpgP2E) were studied by fluorescence techniques, namely: lifetime measurements and acrylamide quenching of the fluorescence of Trp residues. The fluorescence decays of Fpg and its mutant forms were analysed by the maximum-entropy method and lifetime distributions in the range 200 ps to 9 ns were obtained. The lifetime distribution profiles of FpgK57G, FpgP2G and FpgP2E are different from that of wild-type Fpg. Both dynamic and static quenching by acrylamide were observed for all the proteins. At 20 degrees C, the bimolecular collisional quenching rate constant of the FpgP2E fluorescence by acrylamide was only 0.8 M(-1) s(-1) as compared to about 1.4 M(-1) s(-1) for the three other proteins. At 6 degrees C, all the spectroscopic properties of these four proteins are about the same. The analysis of experimental data demonstrates that all three mutations induce a structural reorganization of the Fpg protein. However, only the P2E mutation lead to a reduced accessibility of some Trp residues to acrylamide quenching. It is conclude...Continue Reading

Citations

Oct 6, 2000·Annual Review of Microbiology·M Riley, M H Serres
May 8, 2010·Human Reproduction·Luke SimonSheena E M Lewis
Apr 1, 2000·The Journal of Biological Chemistry·O M Sidorkina, J Laval
Mar 26, 2002·The Journal of Biological Chemistry·Rotem GilboaGil Shoham

Related Concepts

DNA-formamidopyrimidine glycosylase, E coli
Acrylamides
Bacterial Proteins
Alkalescens-Dispar Group
Nucleosidases
T4 DNA Ligase
Protein Conformation
Fluorescence Spectroscopy
PMS-Tryptophan
Point Mutation

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