PURIFICATION AND CHARACTERIZATION OF THE CONSTITUTIVE NITRIC-OXIDE SYNTHASE FROM HUMAN PLACENTA
PURIFICATION AND CHARACTERIZATION OF THE CONSTITUTIVE NITRIC-OXIDE SYNTHASE FROM HUMAN PLACENTA
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DOI：
10.1006/abbi.1994.1232
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发表时间：
1994-06-01
影响因子：
3.9
通讯作者：
CHARLES, IG
中科院分区：
文献类型：
--
作者：
GARVEY, EP;TUTTLE, JV;CHARLES, IG
在人胎盘组织中检测到人内皮型一氧化氮合酶（NOS）mRNA。相反，在胎盘中未检测到人神经元型NOS或人诱导型NOS的mRNA。随后，从胎盘提取物中纯化NOS超过3800倍至大于80%的均一性。通过[I-125]钙调素结合十二烷基硫酸钠-聚丙烯酰胺凝胶中的蛋白质，鉴定出表观分子量为135 kDa的单一条带，这与内皮NOS的预测大小一致。此外，从这个135-kDa的蛋白质衍生的8个内部肽的序列是相同的人内皮NOS的公开序列。正如所有组成型NOS同工酶所示，纯化的NOS绝对依赖于钙和钙调素。NOS也从人脐静脉内皮细胞纯化，并在类似的动力学参数和依赖于钙和钙调素的基础上，似乎是相同的纯化的胎盘NOS。总之，这些数据表明胎盘NOS是来自内皮组织的组成型NOS同工酶。(C)1994年出版社出版。
Human endothelial nitric oxide synthase (NOS) mRNA was detected in human placenta. In contrast, mRNAs for human neuronal NOS or for human inducible NOS were not detected in placenta. Subsequently, NOS was purified over 3800-fold from placental extract to greater than 80% homogeneity. A single band with an apparent molecular weight of 135 kDa was identified by [I-125] calmodulin binding to proteins in a sodium dodecyl sulfate-polyacrylamide gel, which is consistent with the predicted size of the endothelial NOS. Furthermore, the sequence of eight internal peptides derived from this 135-kDa protein was identical to the published sequence of human endothelial NOS. As has been shown for all constitutive NOS isozymes, the purified NOS was absolutely dependent on calcium and calmodulin. NOS was also purified from human umbilical vein endothelial cells and, on the basis of similar kinetic parameters and dependence upon calcium and calmodulin, appeared to be the same as the purified placental NOS. Together, these data indicate that the placental NOS is the constitutive NOS isozyme from endothelial tissue. (C) 1994 Academic Press, Inc.