Catalytic mechanism of the haloalkane dehalogenase LinB from
Sphingomonas paucimobilis UT26

J 2003

Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26

PROKOP, Zbyněk, Marta MONINCOVÁ, Radka CHALOUPKOVÁ, Martin KLVAŇA, Yuji NAGATA et. al.

Základní údaje

Originální název

Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26

Autoři

PROKOP, Zbyněk (203 Česká republika, garant), Marta MONINCOVÁ (203 Česká republika), Radka CHALOUPKOVÁ (203 Česká republika), Martin KLVAŇA (203 Česká republika), Yuji NAGATA (392 Japonsko), Dick B. JANSSEN (528 Nizozemské království) a Jiří DAMBORSKÝ (203 Česká republika)

Vydání

Journal of Biological Chemistry, 2003, 0021-9258

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: 6.482

Kód RIV

RIV/00216224:14310/03:00009155

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000186452300008

Klíčová slova anglicky

ENZYME; KINETICS; MECHANISM; LINB; HALOALKANE DEHALOGENASE

Štítky

Enzyme, haloalkane dehalogenase, kinetics, LINB, mechanism

Změněno: 19. 3. 2010 11:05, prof. Mgr. Jiří Damborský, Dr.

Anotace

V originále

Haloalkane dehalogenases are bacterial enzymes capable of carbon-halogen bond cleavage in halogenated compounds. To obtain insight in the mechanism of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB), we studied the steady-state and pre-steady-state kinetics of the conversion of the substrates 1-chlorohexane, chlorocyclohexane and bromocyclohexane. The results lead to a proposal of a minimal kinetic mechanism consisting of three main steps: (i) substrate binding, (ii) cleavage of the carbon-halogen bond with simultaneous formation of an alkyl-enzyme intermediate and (iii) hydrolysis of the alkyl-enzyme intermediate. Release of both products, halide and alcohol, is a fast process that was not included in reaction mechanism as a distinct step. Comparison of the kinetic mechanism of LinB with that of haloalkane dehalogenase DhlA from Xantobacter autotrophicus GJ10 and the haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 shows that the overall mechanisms are similar. The main difference is in the rate-limiting step, which is hydrolysis of the alkyl-enzyme intermediate in LinB, halide release in DhlA, and liberation of an alcohol in DhaA. The occurrence of different rate-limiting steps for three enzymes that belong to the same protein family indicates that extrapolation of this important catalytic property from one enzyme to another can be misleading even for evolutionary closely related proteins. The differences in the rate-limiting step were related to: (i) a number and size of the entrance tunnels, (ii) protein flexibility and (iii) composition of the halide-stabilizing active site residues based on comparison of protein structures.

Návaznosti

LN00A016, projekt VaV

Název: BIOMOLEKULÁRNÍ CENTRUM

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

Citovat

PROKOP, Zbyněk, Marta MONINCOVÁ, Radka CHALOUPKOVÁ, Martin KLVAŇA, Yuji NAGATA, Dick B. JANSSEN a Jiří DAMBORSKÝ. Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26. Journal of Biological Chemistry. 2003, roč. 278, č. 46, s. 45094-45100. ISSN 0021-9258.

@article{490382,
   author = {Prokop, Zbyněk and Monincová, Marta and Chaloupková, Radka and Klvaňa, Martin and Nagata, Yuji and Janssen, Dick B. and Damborský, Jiří},
   article_number = {46},
   keywords = {ENZYME; KINETICS; MECHANISM; LINB; HALOALKANE DEHALOGENASE},
   language = {eng},
   issn = {0021-9258},
   journal = {Journal of Biological Chemistry},
   title = {Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26},
   url = {http://ncbr.chemi.muni.cz/~jiri/abstracts/jbc03b.html},
   volume = {278},
   year = {2003}
}

TY  - JOUR
ID  - 490382
AU  - Prokop, Zbyněk - Monincová, Marta - Chaloupková, Radka - Klvaňa, Martin - Nagata, Yuji - Janssen, Dick B. - Damborský, Jiří
PY  - 2003
TI  - Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26
JF  - Journal of Biological Chemistry
VL  - 278
IS  - 46
SP  - 45094-45100
EP  - 45094-45100
SN  - 00219258
KW  - ENZYME
KW  - KINETICS
KW  - MECHANISM
KW  - LINB
KW  - HALOALKANE DEHALOGENASE
UR  - http://ncbr.chemi.muni.cz/~jiri/abstracts/jbc03b.html
N2  - Haloalkane dehalogenases are bacterial enzymes capable of carbon-halogen bond cleavage in halogenated compounds. To obtain insight in the mechanism of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB), we studied the steady-state and pre-steady-state kinetics of the conversion of the substrates 1-chlorohexane, chlorocyclohexane and bromocyclohexane. The results lead to a proposal of a minimal kinetic mechanism consisting of three main steps: (i) substrate binding, (ii) cleavage of the carbon-halogen bond with simultaneous formation of an alkyl-enzyme intermediate and (iii) hydrolysis of the alkyl-enzyme intermediate. Release of both products, halide and alcohol, is a fast process that was not included in reaction mechanism as a distinct step. Comparison of the kinetic mechanism of LinB with that of haloalkane dehalogenase DhlA from Xantobacter autotrophicus GJ10 and the haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 shows that the overall mechanisms are similar. The main difference is in the rate-limiting step, which is hydrolysis of the alkyl-enzyme intermediate in LinB, halide release in DhlA, and liberation of an alcohol in DhaA. The occurrence of different rate-limiting steps for three enzymes that belong to the same protein family indicates that extrapolation of this important catalytic property from one enzyme to another can be misleading even for evolutionary closely related proteins. The differences in the rate-limiting step were related to: (i) a number and size of the entrance tunnels, (ii) protein flexibility and (iii) composition of the halide-stabilizing active site residues based on comparison of protein structures.
ER  -

PROKOP, Zbyněk, Marta MONINCOVÁ, Radka CHALOUPKOVÁ, Martin KLVAŇA, Yuji NAGATA, Dick B. JANSSEN a Jiří DAMBORSKÝ. Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26. \textit{Journal of Biological Chemistry}. 2003, roč.~278, č.~46, s.~45094-45100. ISSN~0021-9258.

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