Purification and properties of calf liver gamma-butyrobetaine hydroxylase.
Purification and properties of calf liver gamma-butyrobetaine hydroxylase.
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小牛肝γ-丁甜菜碱羟化酶的纯化和性质。
DOI：
10.1016/0003-9861(81)90374-x
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发表时间：
1981
影响因子：
3.9
通讯作者：
Englard,S
中科院分区：
文献类型：
--
作者：
Kondo,A;Blanchard,JS;Englard,S
牛肝γ-丁酰甜菜碱羟化酶经DEAE、凝胶渗透和羟基磷灰石层析纯化约400倍。均质酶是46，000-道尔顿亚基的二聚体。测定了底物和辅因子的Km值以及抗坏血酸和过氧化氢酶的表观活化常数。通过一些二价金属对酶的抑制支持巯基在金属结合中的功能。已获得抗该酶的抗体，它不与假单胞菌属菌株的均相γ-丁酰甜菜碱羟化酶发生交叉反应。该抗体与Sepharose 4 B偶联，已用于一步纯化小牛肝羟化酶350倍。
Calf liver γ-butyrobetaine hydroxylase has been purified some 400-fold by DEAE, gel permeation, and hydroxylapatite chromatography. The homogeneous enzyme is a dimer of 46,000-dalton subunits. TheKmvalues for substrates and cofactors and the apparent activation constants for ascorbate and catalase have been determined. Inhibition of the enzyme by a number of divalent metals supports the function of sulfhydryl groups in metal binding. An antibody to the enzyme has been obtained; this does not cross-react with homogeneous γ-butyrobetaine hydroxylase from aPseudomonasstrain. The antibody, coupled to Sepharose 4B, has been used to purify the calf liver hydroxylase 350-fold in one step.