Purification and kinetic studies of adenine phosphoribosyltransferase from human erythrocytes

Satish K. Srivastava; Ernest Beutler

doi:10.1016/0003-9861(71)90506-6

Purification and kinetic studies of adenine phosphoribosyltransferase from human erythrocytes

Satish K. Srivastava
, Ernest Beutler

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

Adenine phosphoribosyltransferase was purified 2200-fold from human erythrocytes. The purification steps involved (NH₄)₂SO₄ fractionation followed by Sephadex-G75 column chromatography, DEAE-Sephadex chromatography, and (NH₄)₂SO₄ precipitation. Partially purified enzyme was stabilized by 0.3 m (NH₄)₂SO₄, which also protected the enzyme from heat denaturation. The enzyme was partially inhibited by sodium ions which were partially antagonized by magnesium. Divalent cation is an absolute requirement for high enzyme activity. Ca²⁺, Mn²⁺, and Mg²⁺ supported high initial rates and Hg²⁺ completely inhibited the enzyme. The kinetic data with initial velocity and the product inhibition studies are consistent with a mechanism in which the synthesis of nucleotide proceeds as an ordered sequential reaction.

Original language	English (US)
Pages (from-to)	426-434
Number of pages	9
Journal	Archives of Biochemistry and Biophysics
Volume	142
Issue number	2
DOIs	https://doi.org/10.1016/0003-9861(71)90506-6
State	Published - Feb 1971
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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title = "Purification and kinetic studies of adenine phosphoribosyltransferase from human erythrocytes",

abstract = "Adenine phosphoribosyltransferase was purified 2200-fold from human erythrocytes. The purification steps involved (NH4)2SO4 fractionation followed by Sephadex-G75 column chromatography, DEAE-Sephadex chromatography, and (NH4)2SO4 precipitation. Partially purified enzyme was stabilized by 0.3 m (NH4)2SO4, which also protected the enzyme from heat denaturation. The enzyme was partially inhibited by sodium ions which were partially antagonized by magnesium. Divalent cation is an absolute requirement for high enzyme activity. Ca2+, Mn2+, and Mg2+ supported high initial rates and Hg2+ completely inhibited the enzyme. The kinetic data with initial velocity and the product inhibition studies are consistent with a mechanism in which the synthesis of nucleotide proceeds as an ordered sequential reaction.",

author = "Srivastava, \{Satish K.\} and Ernest Beutler",

note = "Funding Information: in part by Grant Institute, NIH. Funding Information: 1 This work was supported 07449 from the National Heart",

year = "1971",

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doi = "10.1016/0003-9861(71)90506-6",

language = "English (US)",

volume = "142",

pages = "426--434",

journal = "Archives of Biochemistry and Biophysics",

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T1 - Purification and kinetic studies of adenine phosphoribosyltransferase from human erythrocytes

AU - Srivastava, Satish K.

AU - Beutler, Ernest

N1 - Funding Information: in part by Grant Institute, NIH. Funding Information: 1 This work was supported 07449 from the National Heart

PY - 1971/2

Y1 - 1971/2

N2 - Adenine phosphoribosyltransferase was purified 2200-fold from human erythrocytes. The purification steps involved (NH4)2SO4 fractionation followed by Sephadex-G75 column chromatography, DEAE-Sephadex chromatography, and (NH4)2SO4 precipitation. Partially purified enzyme was stabilized by 0.3 m (NH4)2SO4, which also protected the enzyme from heat denaturation. The enzyme was partially inhibited by sodium ions which were partially antagonized by magnesium. Divalent cation is an absolute requirement for high enzyme activity. Ca2+, Mn2+, and Mg2+ supported high initial rates and Hg2+ completely inhibited the enzyme. The kinetic data with initial velocity and the product inhibition studies are consistent with a mechanism in which the synthesis of nucleotide proceeds as an ordered sequential reaction.

AB - Adenine phosphoribosyltransferase was purified 2200-fold from human erythrocytes. The purification steps involved (NH4)2SO4 fractionation followed by Sephadex-G75 column chromatography, DEAE-Sephadex chromatography, and (NH4)2SO4 precipitation. Partially purified enzyme was stabilized by 0.3 m (NH4)2SO4, which also protected the enzyme from heat denaturation. The enzyme was partially inhibited by sodium ions which were partially antagonized by magnesium. Divalent cation is an absolute requirement for high enzyme activity. Ca2+, Mn2+, and Mg2+ supported high initial rates and Hg2+ completely inhibited the enzyme. The kinetic data with initial velocity and the product inhibition studies are consistent with a mechanism in which the synthesis of nucleotide proceeds as an ordered sequential reaction.

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JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 2

ER -

Purification and kinetic studies of adenine phosphoribosyltransferase from human erythrocytes

Abstract

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