Purification and properties of ornithine carbamoyltransferase from loggerhead turtle liver.
Purification and properties of ornithine carbamoyltransferase from loggerhead turtle liver.
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蠵龟肝脏鸟氨酸氨基甲酰转移酶的纯化及其性质。
DOI：
10.33549/physiolres.930193
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发表时间：
2002
影响因子：
2.1
通讯作者：
A. Galtieri
中科院分区：
文献类型：
--
作者：
E. Bellocco;C. D. Salvo;G. Laganà;U. Leuzzi;E. Tellone;A. Kotyk;A. Galtieri
从红海龟Caretta Caretta的肝脏中纯化了鸟氨酸氨基甲酰转移酶，采用亲和柱的一步法纯化，过渡态类似物δ - n-（磷酸乙酰基）- l -鸟氨酸（δ - palo）共价结合。该酶被纯化373倍，经非变性和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）鉴定为均相酶。该酶的比活性为224。C. caretta酶的摩尔质量约为112 kDa， SDS-PAGE单谱显示其亚基摩尔质量为39.5 kDa；因此，酶是相同亚基的三聚体。它催化了一个有序的顺序机制，其中氨基甲酰磷酸盐首先结合，然后是l -鸟氨酸。l -鸟氨酸的Michaelis常数为0.858 mM，氨甲酰磷酸的Michaelis常数为0.22 mM，酶-氨甲酰磷酸配合物的解离常数为0.50 mM。
Ornithine carbamoyltransferase has been purified from the liver of the loggerhead turtle Caretta caretta by a single-step procedure using chromatography on an affinity column to which the transition-state analogue, delta-N-(phosphonoacetyl)-L-ornithine (delta-PALO), was covalently bound. The procedure employed yielded an enzyme which was purified 373-fold and was judged to be homogeneous by nondenaturing and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed a specific activity of 224. The molar mass of the C. caretta enzyme was approximately 112 kDa, the single band obtained by SDS-PAGE indicated a subunit molar mass of 39.5 kDa; hence, the enzyme is a trimer of identical subunits. It catalyzes an ordered sequential mechanism in which carbamoyl phosphate binds first, followed by L-ornithine. The Michaelis constants were 0.858 mM for L-ornithine and 0.22 mM for carbamoyl phosphate, the dissociation constant of the enzyme-carbamoyl phosphate complex was 0.50 mM.
通过 pH 研究确定鸟氨酸转氨甲酰酶的底物特异性和质子化状态。
影响因子：
2.9
作者：
Kuo,LC;Herzberg,W;Lipscomb,WN
通讯作者：
Lipscomb,WN
来自粗糙脉孢菌的鸟氨酸转氨甲酰酶：纯化和特性。
影响因子：
3.9
作者：
Bates,M;Weiss,RL;Clarke,S
通讯作者：
Clarke,S
鸟氨酸转氨甲酰酶的 Zn2 调节。
影响因子：
5.6
作者：
Kuo,LC;Caron,C;Lee,S;Herzberg,W
通讯作者：
Herzberg,W