Dataset Information

Deamination of 6-aminodeoxyfutalosine in menaquinone biosynthesis by distantly related enzymes.

ABSTRACT: Proteins of unknown function belonging to cog1816 and cog0402 were characterized. Sav2595 from Steptomyces avermitilis MA-4680, Acel0264 from Acidothermus cellulolyticus 11B, Nis0429 from Nitratiruptor sp. SB155-2 and Dr0824 from Deinococcus radiodurans R1 were cloned, purified, and their substrate profiles determined. These enzymes were previously incorrectly annotated as adenosine deaminases or chlorohydrolases. It was shown here that these enzymes actually deaminate 6-aminodeoxyfutalosine. The deamination of 6-aminodeoxyfutalosine is part of an alternative menaquinone biosynthetic pathway that involves the formation of futalosine. 6-Aminodeoxyfutalosine is deaminated by these enzymes with catalytic efficiencies greater than 10(5) M(-1) s(-1), Km values of 0.9-6.0 ?M, and kcat values of 1.2-8.6 s(-1). Adenosine, 2'-deoxyadenosine, thiomethyladenosine, and S-adenosylhomocysteine are deaminated at least an order of magnitude slower than 6-aminodeoxyfutalosine. The crystal structure of Nis0429 was determined and the substrate, 6-aminodeoxyfutalosine, was positioned in the active site on the basis of the presence of adventitiously bound benzoic acid. In this model, Ser-145 interacts with the carboxylate moiety of the substrate. The structure of Dr0824 was also determined, but a collapsed active site pocket prevented docking of substrates. A computational model of Sav2595 was built on the basis of the crystal structure of adenosine deaminase and substrates were docked. The model predicted a conserved arginine after ?-strand 1 to be partially responsible for the substrate specificity of Sav2595.

SUBMITTER: Goble AM

PROVIDER: S-EPMC3813303 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature

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Deamination of 6-aminodeoxyfutalosine in menaquinone biosynthesis by distantly related enzymes.

Goble Alissa M AM Toro Rafael R Li Xu X Ornelas Argentina A Fan Hao H Eswaramoorthy Subramaniam S Patskovsky Yury Y Hillerich Brandan B Seidel Ron R Sali Andrej A Shoichet Brian K BK Almo Steven C SC Swaminathan Subramanyam S Tanner Martin E ME Raushel Frank M FM

Biochemistry 20130904 37

Proteins of unknown function belonging to cog1816 and cog0402 were characterized. Sav2595 from Steptomyces avermitilis MA-4680, Acel0264 from Acidothermus cellulolyticus 11B, Nis0429 from Nitratiruptor sp. SB155-2 and Dr0824 from Deinococcus radiodurans R1 were cloned, purified, and their substrate profiles determined. These enzymes were previously incorrectly annotated as adenosine deaminases or chlorohydrolases. It was shown here that these enzymes actually deaminate 6-aminodeoxyfutalosine. Th ...[more]

PMID: 23972005

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Dataset Information

Deamination of 6-aminodeoxyfutalosine in menaquinone biosynthesis by distantly related enzymes.

Publications

Deamination of 6-aminodeoxyfutalosine in menaquinone biosynthesis by distantly related enzymes.

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