Exploring the structure and activity of haloalkane dehalogenase
from Sphingomonas paucimobilis UT26: evidence for product and
water mediated inhibition

J 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

URL

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

dehalogenation, Enzyme, inhibition, Protein engineering, x-ray

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

Název: Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

MSM 143100005, záměr

Název: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Citovat

OAKLEY, Aaron, Zbyněk PROKOP, Michal BOHÁČ, Jan KMUNÍČEK, Tomáš JEDLIČKA, Marta MONINCOVÁ, Ivana KUTÁ-SMATANOVÁ, Yuji NAGATA, Jiří DAMBORSKÝ a Matthew WILCE. Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition. Biochemistry. 2002, roč. 41, č. 15, s. 4847-4855. ISSN 0006-2960.

@article{404587,
   author = {Oakley, Aaron and Prokop, Zbyněk and Boháč, Michal and Kmuníček, Jan and Jedlička, Tomáš and Monincová, Marta and KutáandSmatanová, Ivana and Nagata, Yuji and Damborský, Jiří and Wilce, Matthew},
   article_number = {15},
   keywords = {X-RAY; DEHALOGENATION; ENZYME; PROTEIN ENGINEERING; INHIBITION},
   language = {eng},
   issn = {0006-2960},
   journal = {Biochemistry},
   title = {Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition},
   url = {http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/biochem02.html},
   volume = {41},
   year = {2002}
}

TY  - JOUR
ID  - 404587
AU  - Oakley, Aaron - Prokop, Zbyněk - Boháč, Michal - Kmuníček, Jan - Jedlička, Tomáš - Monincová, Marta - Kutá-Smatanová, Ivana - Nagata, Yuji - Damborský, Jiří - Wilce, Matthew
PY  - 2002
TI  - Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition
JF  - Biochemistry
VL  - 41
IS  - 15
SP  - 4847
EP  - 4847
SN  - 00062960
KW  - X-RAY
KW  - DEHALOGENATION
KW  - ENZYME
KW  - PROTEIN ENGINEERING
KW  - INHIBITION
UR  - http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/biochem02.html
N2  - 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.
ER  -

OAKLEY, Aaron, Zbyněk PROKOP, Michal BOHÁČ, Jan KMUNÍČEK, Tomáš JEDLIČKA, Marta MONINCOVÁ, Ivana KUTÁ-SMATANOVÁ, Yuji NAGATA, Jiří DAMBORSKÝ a Matthew WILCE. Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition. \textit{Biochemistry}. 2002, roč.~41, č.~15, s.~4847-4855. ISSN~0006-2960.

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