カツオ肝臓のNAD(P)加水分解酵素-III

阿部 宏喜; 清水 千秋; 松浦 文雄

doi:10.2331/suisan.44.279

NAD (P) Hydrolyzing Enzymes in Skipjack Liver-III

Purification and Properties of Nucleotide Pyrophosphatase

Hiroki ABE, Chiaki SHIMIZU, Fumio MATSUURA

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JOURNALS FREE ACCESS

Volume 44 (1978) Issue 3 Pages 279-284

DOI https://doi.org/10.2331/suisan.44.279

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Released: February 29, 2008 [Advance Publication] Released: - received: September 20, 1977 corrected: - accepted: -
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Date of correction: February 29, 2008 Reason for correction: - Correction: ABSTRACT Details: Wrong : Nucleotide pyrophosphatase was purified from skipjack liver by various types of column chromatography and gel filtration. The enzyme was ultimately purified aout 2, 900-fold. It was homogeneous in polyacrylamide gel electrophoresis, but exhibited a slight alkaline phosphatase activity, sugesting same persistent contamination. It was found by analytical gel filtration and SDS gel electrophoresis that the enzyme has a molecular weight of 86, 000 and is composed of a single polypeptide chain. The isoelectric point was determined to be 4.8 by isoelectrofocusing.
The enzyme showed its maximum activity at around pH9 when NAD or 2′-deoxythymidine-5′-p-nitrophenyl phosphate was used as the substrate. It was to some extent activated by Mg²⁺, but inhibited by other metal ions, such as Mn²⁺, Fe²⁺, Fe³⁺, Ca²⁺, as well as by several chelating agents. The enzyme was inhibited competitively by 5′-AMP and nicotinamide mononucleotide, both of which are reaction products from NAD, and also by other nucleotides. The skipjack enzyme showed a higher affinity and maximum velocity for NADH₂ than for NAD (P). The enzyme hydrolyzed various sugar nucleotides, ATP, ADP, and inorganic pyrophosphate, thus indicating a low substrate specificity.
Right : Nucleotide pyrophosphatase was purified from skipjack liver by various types of column chromatography and gel filtration. The enzyme was ultimately purified aout 2, 900-fold. It was homogeneous in polyacrylamide gel electrophoresis, but exhibited a slight alkaline phosphatase activity, sugesting same persistent contamination. It was found by analytical gel filtration and SDS gel electrophoresis that the enzyme has a molecular weight of 86, 000 and is composed of a single polypeptide chain. The isoelectric point was determined to be 4.8 by isoelectrofocusing.
The enzyme showed its maximum activity at around pH9 when NAD or 2'-deoxythymidine-5'-p-nitrophenyl phosphate was used as the substrate. It was to some extent activated by Mg²⁺, but inhibited by other metal ions, such as Mn²⁺, Fe²⁺, Fe³⁺, Ca²⁺, as well as by several chelating agents. The enzyme was inhibited competitively by 5'-AMP and nicotinamide mononucleotide, both of which are reaction products from NAD, and also by other nucleotides. The skipjack enzyme showed a higher affinity and maximum velocity for NADH₂ than for NAD (P). The enzyme hydrolyzed various sugar nucleotides, ATP, ADP, and inorganic pyrophosphate, thus indicating a low substrate specificity.

Date of correction: February 29, 2008 Reason for correction: - Correction: PDF FILE Details: -

Abstract

Nucleotide pyrophosphatase was purified from skipjack liver by various types of column chromatography and gel filtration. The enzyme was ultimately purified aout 2, 900-fold. It was homogeneous in polyacrylamide gel electrophoresis, but exhibited a slight alkaline phosphatase activity, sugesting same persistent contamination. It was found by analytical gel filtration and SDS gel electrophoresis that the enzyme has a molecular weight of 86, 000 and is composed of a single polypeptide chain. The isoelectric point was determined to be 4.8 by isoelectrofocusing.
The enzyme showed its maximum activity at around pH9 when NAD or 2'-deoxythymidine-5'-p-nitrophenyl phosphate was used as the substrate. It was to some extent activated by Mg²⁺, but inhibited by other metal ions, such as Mn²⁺, Fe²⁺, Fe³⁺, Ca²⁺, as well as by several chelating agents. The enzyme was inhibited competitively by 5'-AMP and nicotinamide mononucleotide, both of which are reaction products from NAD, and also by other nucleotides. The skipjack enzyme showed a higher affinity and maximum velocity for NADH₂ than for NAD (P). The enzyme hydrolyzed various sugar nucleotides, ATP, ADP, and inorganic pyrophosphate, thus indicating a low substrate specificity.

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