TY - JOUR
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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U2 - 10.1016/0003-9861(71)90506-6
DO - 10.1016/0003-9861(71)90506-6
M3 - Article
C2 - 4323725
AN - SCOPUS:0015011371
SN - 0003-9861
VL - 142
SP - 426
EP - 434
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -