Identification of protein fold and catalytic residues of
g-hexachlorocyclohexane dehydrochlorinase LinA

J 2001

Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA

NAGATA, Yuji; Katsuki MORI; Masamichi TAKAGI; Alexey G. MURZIN; Jiri DAMBORSKY et. al.

Základní údaje

Originální název

Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA

Autoři

NAGATA, Yuji; Katsuki MORI; Masamichi TAKAGI; Alexey G. MURZIN a Jiri DAMBORSKY

Vydání

PROTEINS: Structure, Function, and Genetics, 2001

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

Kód RIV

RIV/00216224:14310/01:00005567

Organizační jednotka

Přírodovědecká fakulta

Změněno: 18. 3. 2002 15:18, prof. Mgr. Jiří Damborský, Dr.

Anotace

V originále

g-Hexachlorocyclohexane dehydrochlorinase (LinA) is a unique dehydrochlorinase that has no homologous sequence at the amino acid-sequence level, and for which the evolutionary origin is unknown. We here propose that LinA is a member of a novel structural superfamily of enzymes containing scytalone dehydratase, 3-oxo-D5-steroid isomerase, nuclear transport factor-2, and the b-subunit of naphthalene dioxygenase-all known structures with different functions. The catalytic and the active site residues of LinA are predicted based on its homology model. Nine mutants that carry substitutions in the proposed catalytic residues were constructed by site-directed mutagenesis. In addition to these, eight mutants that have a potential to make contact with the substrate were prepared by site-directed mutagenesis. These mutants were expressed in E. coli, and their activities in crude extract were evaluated. Most of the features of the LinA mutants could be explained on the basis of the present LinA model, indicating its validity. We conclude that LinA catalyses the proton abstraction via the catalytic dyad H73-D25 by the similar mechanism as described for scytalone dehydratase. The results suggest that LinA and scytalone dehydratase evolved from a common ancestor. LinA may have evolved from an enzyme showing a dehydratase activity.

Návaznosti

LN00A016, projekt VaV

Název: BIOMOLEKULÁRNÍ CENTRUM

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Biomolekulární centrum

Citovat

NAGATA, Yuji; Katsuki MORI; Masamichi TAKAGI; Alexey G. MURZIN a Jiri DAMBORSKY. Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA. PROTEINS: Structure, Function, and Genetics. 2001, roč. 45, č. 4, s. 471-477.

@article{400031,
   author = {Nagata, Yuji and Mori, Katsuki and Takagi, Masamichi and Murzin, Alexey G. and Damborsky, Jiri},
   article_number = {4},
   language = {eng},
   journal = {PROTEINS: Structure, Function, and Genetics},
   title = {Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA},
   url = {http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/prot01.html},
   volume = {45},
   year = {2001}
}

TY  - JOUR
ID  - 400031
AU  - Nagata, Yuji - Mori, Katsuki - Takagi, Masamichi - Murzin, Alexey G. - Damborsky, Jiri
PY  - 2001
TI  - Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA
JF  - PROTEINS: Structure, Function, and Genetics
VL  - 45
IS  - 4
SP  - 471
EP  - 471
UR  - http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/prot01.html
N2  - g-Hexachlorocyclohexane dehydrochlorinase (LinA) is a unique dehydrochlorinase that has no homologous sequence at the amino acid-sequence level, and for which the evolutionary origin is unknown. We here propose that LinA is a member of a novel structural superfamily of enzymes containing scytalone dehydratase, 3-oxo-D5-steroid isomerase, nuclear transport factor-2, and the b-subunit of naphthalene dioxygenase-all known structures with different functions. The catalytic and the active site residues of LinA are predicted based on its homology model. Nine mutants that carry substitutions in the proposed catalytic residues were constructed by site-directed mutagenesis. In addition to these, eight mutants that have a potential to make contact with the substrate were prepared by site-directed mutagenesis. These mutants were expressed in E. coli, and their activities in crude extract were evaluated. Most of the features of the LinA mutants could be explained on the basis of the present LinA model, indicating its validity. We conclude that LinA catalyses the proton abstraction via the catalytic dyad H73-D25 by the similar mechanism as described for scytalone dehydratase. The results suggest that LinA and scytalone dehydratase evolved from a common ancestor. LinA may have evolved from an enzyme showing a dehydratase activity.
ER  -

NAGATA, Yuji; Katsuki MORI; Masamichi TAKAGI; Alexey G. MURZIN a Jiri DAMBORSKY. Identification of protein fold and catalytic residues of g-hexachlorocyclohexane dehydrochlorinase LinA. \textit{PROTEINS: Structure, Function, and Genetics}. 2001, roč.~45, č.~4, s.~471-477.

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