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Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus

Nord, Johan; Kiefer, Martin; Adolph, Hans-Werner; Zeppezauer, Michael M and Nyman, Per-Olof LU (2000) In FEBS Letters 472(2-3). p.312-316
Abstract
Transient kinetics of the equine infectious anemia virus deoxyuridine 5'-triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one-step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis.
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
C-terminus, Deoxyuridine 5'-triphosphate nucleotide hydrolase, Deoxyuridine, Equine infectious anemia virus, Ligand binding, Pre-steady-state kinetics, Rate constant
in
FEBS Letters
volume
472
issue
2-3
pages
312 - 316
publisher
Wiley-Blackwell
external identifiers
  • scopus:0034724658
ISSN
1873-3468
DOI
10.1016/S0014-5793(00)01453-8
language
English
LU publication?
yes
id
dd0ffb89-7225-49b5-b975-8f5025bf8efe (old id 125130)
date added to LUP
2007-07-09 08:24:24
date last changed
2017-08-06 04:31:10
@article{dd0ffb89-7225-49b5-b975-8f5025bf8efe,
  abstract     = {Transient kinetics of the equine infectious anemia virus deoxyuridine 5'-triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one-step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis.},
  author       = {Nord, Johan and Kiefer, Martin and Adolph, Hans-Werner and Zeppezauer, Michael M and Nyman, Per-Olof},
  issn         = {1873-3468},
  keyword      = {C-terminus,Deoxyuridine 5'-triphosphate nucleotide hydrolase,Deoxyuridine,Equine infectious anemia virus,Ligand binding,Pre-steady-state kinetics,Rate constant},
  language     = {eng},
  number       = {2-3},
  pages        = {312--316},
  publisher    = {Wiley-Blackwell},
  series       = {FEBS Letters},
  title        = {Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus},
  url          = {http://dx.doi.org/10.1016/S0014-5793(00)01453-8},
  volume       = {472},
  year         = {2000},
}