PROKOP, Zbyněk, Marta MONINCOVÁ, Radka CHALOUPKOVÁ, Martin KLVAŇA, Yuji NAGATA, Dick B. JANSSEN and Jiří DAMBORSKÝ. Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26. Journal of Biological Chemistry. 2003, vol. 278, No 46, p. 45094-45100. ISSN 0021-9258.
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Basic information
Original name Catalytic mechanism of the haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26
Authors PROKOP, Zbyněk (203 Czech Republic, guarantor), Marta MONINCOVÁ (203 Czech Republic), Radka CHALOUPKOVÁ (203 Czech Republic), Martin KLVAŇA (203 Czech Republic), Yuji NAGATA (392 Japan), Dick B. JANSSEN (528 Netherlands) and Jiří DAMBORSKÝ (203 Czech Republic).
Edition Journal of Biological Chemistry, 2003, 0021-9258.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10600 1.6 Biological sciences
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 6.482
RIV identification code RIV/00216224:14310/03:00009155
Organization unit Faculty of Science
UT WoS 000186452300008
Keywords in English ENZYME; KINETICS; MECHANISM; LINB; HALOALKANE DEHALOGENASE
Tags Enzyme, haloalkane dehalogenase, kinetics, LINB, mechanism
Changed by Changed by: prof. Mgr. Jiří Damborský, Dr., učo 1441. Changed: 19/3/2010 11:05.
Abstract
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.
Links
LN00A016, research and development projectName: BIOMOLEKULÁRNÍ CENTRUM
Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Center
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