2002
Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition
OAKLEY, Aaron, Zbyněk PROKOP, Michal BOHÁČ, Jan KMUNÍČEK, Tomáš JEDLIČKA et. al.Základní údaje
Originální název
Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition
Autoři
OAKLEY, Aaron (36 Austrálie), Zbyněk PROKOP (203 Česká republika), Michal BOHÁČ (203 Česká republika), Jan KMUNÍČEK (203 Česká republika), Tomáš JEDLIČKA (203 Česká republika), Marta MONINCOVÁ (203 Česká republika), Ivana KUTÁ-SMATANOVÁ (203 Česká republika), Yuji NAGATA (392 Japonsko), Jiří DAMBORSKÝ (203 Česká republika, garant) a Matthew WILCE (36 Austrálie)
Vydání
Biochemistry, 2002, 0006-2960
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
Impakt faktor
Impact factor: 4.064
Kód RIV
RIV/00216224:14310/02:00006262
Organizační jednotka
Přírodovědecká fakulta
UT WoS
000175012900011
Klíčová slova anglicky
X-RAY; DEHALOGENATION; ENZYME; PROTEIN ENGINEERING; INHIBITION
Štítky
Změněno: 19. 3. 2010 10:54, prof. Mgr. Jiří Damborský, Dr.
Anotace
V originále
The hydrolysis of haloalkanes to their corresponding alcohols and inorganic halides is catalysed by a/b-hydrolases called haloalkane dehalogenases. The study of haloalkane dehalogenases is vital for the development of these enzymes if they are to be utilized for bioremediation of organohalide-contaminated industrial waste. We report the kinetic and structural analysis of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB) in complex with each of 1,2-dichloroethane and 1,2-dichloropropane and the reaction product of 1-chlorobutane turnover. Activity studies showed very weak, but detectable activity of LinB with 1,2-dichloroethane (0.012 nmol.s-1.mg-1 of enzyme) and 1,2-dichloropropane (0.027 nmol.s-1.mg-1 of enzyme). These activities are much weaker compared, for example, to activity of LinB with 1-chlorobutane (68.169 nmol.s-1.mg-1 of enzyme). Inhibition analysis reveals that both 1,2-dichloroethane and 1,2-dichloropropane act as simple competitive inhibitors of the substrate 1-chlorobutane and that 1,2-dichloroethane binds to LinB with lower affinity than 1,2-dichloropropane. Docking calculations on the enzyme in the absence of active site water molecules and halide ions confirms that these compounds could bind productively. However, when these moieties were included in the calculations, they bound in the manner similar to that observed in the crystal structure. These data provide an explanation for the low activity of LinB with small, chlorinated alkanes and show the importance of active site water molecules and reaction products in molecular docking.
Návaznosti
ME 276, projekt VaV |
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MSM 143100005, záměr |
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