Michel G, Chauvet J, Acher R
Laboratory of Biological Chemistry, University of Paris VI 96, France.
Neuropeptides. 1991 Dec;20(4):211-6. doi: 10.1016/0143-4179(91)90010-g.
Human MSEL-neurophysin has been dissected into two halves by endopeptidase Lys-C, taking advantage of a peculiar Lys59-Ala60 bond. Two sub-domains, N-terminal (1-59) and C-terminal (60-93), have been separated. These sub-domains have been purified by reverse-phase high pressure liquid chromatography and identified by their N-terminal sequences. The N-terminal fragment comprises two chains 1-18 and 19-59, because of the presence of a second lysine residue in position 18, whereas the C-terminal fragment (60-93) is a single chain. Hormone-binding experiments have been carried out using vasopressin or vasopressinyl-Gly-Lys-Arg and testing the ability of the hormone-neurophysin complex to precipitate at pH 3.9 with 10% NaCl. The N-terminal sub-domain precipitates in presence of vasopressin in the same way as native neurophysin whereas the C-terminal sub-domain does not. It can be concluded that the hormone-binding site is located in the 1-59 region of neurophysin.
利用特殊的赖氨酸59 - 丙氨酸60键，人MSEL - 神经垂体素被内肽酶Lys - C切割成两半。两个亚结构域，即N端（1 - 59）和C端（60 - 93）已被分离。这些亚结构域通过反相高压液相色谱法进行了纯化，并通过其N端序列进行了鉴定。由于在第18位存在第二个赖氨酸残基，N端片段包含两条链1 - 18和19 - 59，而C端片段（60 - 93）是单链。使用加压素或加压素基 - 甘氨酸 - 赖氨酸 - 精氨酸进行了激素结合实验，并测试了激素 - 神经垂体素复合物在pH 3.9和10%氯化钠存在下沉淀的能力。N端亚结构域在加压素存在下与天然神经垂体素以相同方式沉淀，而C端亚结构域则不会。可以得出结论，激素结合位点位于神经垂体素的1 - 59区域。
