The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F06%3A00017450" target="_blank" >RIV/00216224:14310/06:00017450 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/07:00022383

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution

Original language description

Haloalkane dehalogenase DhmA from Mycobacterium avium N85 showed poor expression and low stability when produced in Escherichia coli. Here we present expression DhmA in newly constructed pK4RP rhodococcal expression system in a soluble and stable form. Site-directed mutagenesis was used for the identification of a catalytic pentad, which makes up the reaction machinery of all currently known haloalkane dehalogenases. The putative catalytic triad Asp123, His279, Asp250 and the first halide-stabilizing residue Trp124 were deduced from sequence comparisons. The second stabilizing residue Trp164 was predicted from a homology model. Five point mutants in the catalytic pentad were constructed, tested for activity and were found inactive. A two-step reactionmechanism was proposed for DhmA. Evolution of different types of catalytic pentads and molecular adaptation towards the synthetic substrate 1,2-dichloroethane within the protein family is discussed.

Czech name

Identifikace katalytických pentád v haloalkán dehalogenáze DhmA y bakterie Mycobacterium avium N85:reakční mechanismus a molekulární evoluce.

Czech description

Haloalkán dehalogenáza DhmA z bakterie Mycobacterium avium N85 vykazovala nizkou stabilitu a spatnou expresi, kdyz byla vložena do Escherichia coli. v článku je prezentována exprese DhmA v nově konstruovaný rhodokokální expresní systém pK4RP,který byl stabilní........

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

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Continuities

Z - Vyzkumny zamer (s odkazem do CEZ)

Publication year

2006

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

JOURNAL OF STRUCTURAL BIOLOGY

ISSN

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e-ISSN

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Volume of the periodical

1/2006

Issue of the periodical within the volume

1

Country of publishing house

US - UNITED STATES

Number of pages

9

Pages from-to

9999

UT code for WoS article

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EID of the result in the Scopus database

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