Purification and properties of 5'-nucleotidase from the membrane of Vibrio costicola, a moderately halophilic bacterium.

Journal of Bacteriology
C Bengis-Garber, D J Kushner

Abstract

Two different Mg2+-dependent adenosine 5'-triphosphate-hydrolyzing activities were detected in membranes of Vibrio costicola, a novel 5'-nucleotidase and an N,N'-dicyclohexylcarbodiimide-sensitive adenosine triphosphatase. The former and the latter had different requirements for Mg2+ and were selectively assayed in the membranes by using, respectively, 20 and 2 mM Mg2+. The two enzymes were extracted with a combination of Triton X-100 and octylglucoside, separated on a diethylaminoethyl cellulose column, and purified on glycerol gradients. The purified 5'-nucleotidase consisted of one major polypeptide of 70,000 daltons when analyzed on polyacrylamide gels in the presence of sodium dodecyl sulfate. The purified 5'-nucleotidase was similar in substrate specificities, divalent cation specificities, and pH profiles to the membrane-bound N,N'-dicyclohexylcarbodiimide-insensitive nucleotide-phosphohydrolyzing activity. The enzyme hydrolyzed nucleoside 5'-tri, 5'-di, and 5'-monophosphates at comparable rates. Inorganic pyrophosphate, p-nitrophenyl phosphate, glucose 6-phosphate, beta-glycerophosphate, adenosine 5'-diphosphate glucose, adenosine 3'-monophosphate, and cyclic adenosine 3',5'-monophosphate were not hydrolyzed, either im ...Continue Reading

References

Aug 1, 1977·Canadian Journal of Biochemistry·E de Médicis, B Rossignol
May 1, 1977·Journal of Bacteriology·D B ShindlerD J Kushner
Feb 1, 1975·Journal of Bacteriology·E Yagil, I R Beacham
Sep 1, 1969·Journal of Bacteriology·J ThompsonF C Happold
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Dec 17, 1962·Biochimica Et Biophysica Acta·J H CHRISTIAN, J A WALTHO
Oct 1, 1963·Journal of Bacteriology·J KOHN, J L REIS

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Citations

Feb 1, 1989·Journal of Bioenergetics and Biomembranes·N J Russell
Jan 5, 2017·Extremophiles : Life Under Extreme Conditions·Taka-Aki KuribayashiYoshihiro Sambongi
Jul 1, 1988·Applied and Environmental Microbiology·J H PaulW H Jeffrey
Jun 10, 1998·Microbiology and Molecular Biology Reviews : MMBR·A VentosaA Oren
Apr 14, 1987·Biochemical and Biophysical Research Communications·Y SakaiT Tsuchiya

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