Any of several lysines can react with 5'-isothiocyanatofluorescein to inactivate sodium and potassium ion activated adenosinetriphosphatase.
Any of several lysines can react with 5'-isothiocyanatofluorescein to inactivate sodium and potassium ion activated adenosinetriphosphatase.
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几种赖氨酸中的任何一种都可以与 5-异硫氰酸荧光素反应，使钠离子和钾离子激活的腺苷三磷酸酶失活。
作者：
Xu,KY
Kai-yuan Xu Department of Chemistry，D-006，University of加州圣地亚哥，拉霍亚，加州92093接收日期：1988年10月28日;修订后的手稿接收日期：1989年2月8日摘要：反应化学计量的测定表明，一个S '-异硫氰酸荧光素可以抑制一个分子的钠和钾离子激活的腺苷三磷酸酶，这与早先测定的化学计量学一致。然而，当修饰的酶用胰蛋白酶消化时，产生几种不同的修饰肽。其中一种肽已被鉴定为HLLVMK（硫脲基荧光素）GAPER，通过使用特异性免疫吸附剂。修饰的赖氨酸是（Na++ K+）-ATP酶的多肽的氨基酸序列中的赖氨酸501。这种肽以前已从这种寄生虫中分离出来[法利，R.一、特兰角，澳-地M.，卡里利角T.，Hawke，D.，& Shively，J. E.（1984）J.Biol.Chem.259，9532-9535]。其他特异性修饰的肽已被纯化并通过氨基酸测序鉴定。它们的序列鉴定出a多肽中的赖氨酸480和赖氨酸766是在对ATP的加入敏感的反应中被S ′-异硫氰酸荧光素修饰的氨基酸，并负责使酶失活。（Skou，1957）是负责钠和钾通过所有动物细胞质膜的偶联、主动转运的酶（Kyte，1981）。该酶由一个α亚基和一个β亚基组成。重组DNA技术的最新进展使这两种多肽的一级结构的测定变得容易。a亚基，由多肽组成
Kai-yuan Xu Department of Chemistry, D-006, University of California at San Diego, La Jolla, California 92093 Received October 28, 1988; Revised Manuscript Received February 8, 1989 abstract: Determinations of reaction stoichiometry demonstrate that the covalent incorporation of one molecule of S'-isothiocyanatofluorescein can inactivate one molecule of sodium and potassium ion activated adenosinetriphosphatase in agreement with earlier determination of this stoichiometry. Several different modified peptides are produced, however, when the modified enzyme is digested with trypsin. One of these peptides has been identified as HLLVMK (thioureidylfluorescein) GAPER by use of a specific immu-noadsorbent. The modified lysine is lysine 501 in the amino acid sequence of the a polypeptide of (Na++ K+)-ATPase. This peptide has been previously isolated from such digests [Farley, R. A., Tran, C. M., Carilli, C. T., Hawke, D., & Shively, J. E.(1984) J. Biol. Chem. 259, 9532-9535]. The other specifically modified peptideshave been purified and identified by amino acid sequencing. Their sequences identify lysine 480 and lysine 766 from the a polypeptide as amino acids modified by S'-isothiocyanatofluorescein in reactions sensitive to the addition of ATP and responsible for inactivation of theenzyme.Sodium and potassium ion activated adenosinetriphosphatase [(Na++ K+)-ATPase] 1 (Skou, 1957) is the enzyme responsible for the coupled, active transport of sodiumand potassium across the plasma membranes of all animal cells (Kyte, 1981). The enzyme is composed of one a subunit and one ß subunit. Recent advances in recombinant DNA technology have fa-cilitated the determination of the primary structure of the two polypeptides. The a subunit, composed from a polypeptide