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

J 2007

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

PAVLOVÁ, Martina; Martin KLVAŇA; Andrea JESENSKÁ; Zbyněk PROKOP; Hana KONEČNÁ et al.

Základní údaje

Originální název

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

Název česky

Identifikace katalytickych pentad v haloalkán dehalogenaze DhmA z kmene Mycobacterium avium N85: Reakční mechanismus a molekulární evoluce.

Autoři

PAVLOVÁ, Martina; Martin KLVAŇA; Andrea JESENSKÁ; Zbyněk PROKOP; Hana KONEČNÁ; Yukari SATO; M. TSUDA; Yuji NAGATA a Jiří DAMBORSKÝ

Vydání

Journal of Structural Biology, 2007, 1047-8477

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.677

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/07:00022383

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000243951800010

Klíčová slova anglicky

Catalytic Pentad; Haloalkane Dehalogenase; DhmA Mycobacterium avium N85

Štítky

Catalytic Pentad, DhmA Mycobacterium avium N85, haloalkane dehalogenase

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 22. 3. 2010 09:10, prof. Mgr. Jiří Damborský, Dr.

Anotace

ORIG CZ

V originále

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 reaction mechanism 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.

Česky

v clanku jsou popsany experimenty s Haloalkan dehalogenázou DhmA z bakterie Mycobacterium avium N85.

Návaznosti

MSM0021622413, záměr

Název: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Proteiny v metabolismu a při interakci organismů s prostředím

Citovat

PAVLOVÁ, Martina; Martin KLVAŇA; Andrea JESENSKÁ; Zbyněk PROKOP; Hana KONEČNÁ; Yukari SATO; M. TSUDA; Yuji NAGATA a Jiří DAMBORSKÝ. The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution. Journal of Structural Biology. 2007, roč. 157, č. 2, s. 384-392. ISSN 1047-8477.

@article{721200,
   author = {Pavlová, Martina and Klvaňa, Martin and Jesenská, Andrea and Prokop, Zbyněk and Konečná, Hana and Sato, Yukari and Tsuda, M. and Nagata, Yuji and Damborský, Jiří},
   article_number = {2},
   keywords = {Catalytic Pentad; Haloalkane Dehalogenase; DhmA Mycobacterium avium N85},
   language = {eng},
   issn = {1047-8477},
   journal = {Journal of Structural Biology},
   title = {The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution},
   url = {http://loschmidt.chemi.muni.cz/peg/abstracts/jsb07.html},
   volume = {157},
   year = {2007}
}

TY  - JOUR
ID  - 721200
AU  - Pavlová, Martina - Klvaňa, Martin - Jesenská, Andrea - Prokop, Zbyněk - Konečná, Hana - Sato, Yukari - Tsuda, M. - Nagata, Yuji - Damborský, Jiří
PY  - 2007
TI  - The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution
JF  - Journal of Structural Biology
VL  - 157
IS  - 2
SP  - 384-392
EP  - 384-392
SN  - 10478477
KW  - Catalytic Pentad
KW  - Haloalkane Dehalogenase
KW  - DhmA Mycobacterium avium N85
UR  - http://loschmidt.chemi.muni.cz/peg/abstracts/jsb07.html
N2  - 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 reaction mechanism 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.
ER  -

PAVLOVÁ, Martina; Martin KLVAŇA; Andrea JESENSKÁ; Zbyněk PROKOP; Hana KONEČNÁ; Yukari SATO; M. TSUDA; Yuji NAGATA a Jiří DAMBORSKÝ. The Identification of Catalytic Pentad in the Haloalkane Dehalogenase DhmA from Mycobacterium avium N85: Reaction Mechanism and Molecular Evolution. \textit{Journal of Structural Biology}. 2007, roč.~157, č.~2, s.~384-392. ISSN~1047-8477.

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