A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F12%3A00057142" target="_blank" >RIV/00216224:14310/12:00057142 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1074/jbc.M112.377853" target="_blank" >http://dx.doi.org/10.1074/jbc.M112.377853</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1074/jbc.M112.377853" target="_blank" >10.1074/jbc.M112.377853</a>
Alternative languages
Result language
angličtina
Original language name
A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB
Original language description
Many enzymes have buried active sites. The properties of the tunnels connecting the active site with bulk solvent affect ligand binding and unbinding and also the catalytic properties. Here, we investigate ligand passage in the haloalkane dehalogenase enzyme LinB and the effect of replacing leucine by a bulky tryptophan at a tunnel-lining position. Transient kinetic experiments show that the mutation significantly slows down the rate of product release. Moreover, the mechanism of bromide ion release ischanged from a one-step process in the wild type enzyme to a two-step process in the mutant. The rate constant of bromide ion release corresponds to the overall steady-state turnover rate constant, suggesting that product release became the rate-limitingstep of catalysis in the mutant. We explain the experimental findings by investigating the molecular details of the process computationally.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EB - Genetics and molecular biology
OECD FORD branch
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Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2012
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
The Journal of Biological Chemistry
ISSN
0021-9258
e-ISSN
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Volume of the periodical
287
Issue of the periodical within the volume
34
Country of publishing house
US - UNITED STATES
Number of pages
13
Pages from-to
29062-29074
UT code for WoS article
000308074600073
EID of the result in the Scopus database
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