Kirley T L, Wallick E T, Lane L K
Biochem Biophys Res Commun. 1984 Dec 14;125(2):767-73. doi: 10.1016/0006-291x(84)90605-3.
Fluorescein 5'-isothiocyanate has been used to label ouabain sensitive and insensitive (Na,K)-ATPases from lamb and rat kidney, respectively. The labeled enzymes were digested with trypsin to generate soluble peptides, which were purified by high performance liquid chromatography and sequenced on a gas phase sequenator. The sequence of the labeled peptide from both species is His-Leu-Leu-Val-Met-Lys-Gly-Ala-Pro-Glu-Arg. Thus, it appears that the primary structure of the fluorescein 5'-isothiocyanate reactive site, and therefore presumably the ATP binding site, is completely conserved in ouabain sensitive and ouabain insensitive (Na,K)-ATPases.
5'-异硫氰酸荧光素已分别用于标记来自羔羊和大鼠肾脏的哇巴因敏感型和不敏感型(钠,钾)-ATP酶。用胰蛋白酶消化标记的酶以产生可溶性肽,这些肽通过高效液相色谱法纯化并在气相测序仪上测序。两种物种的标记肽序列均为His-Leu-Leu-Val-Met-Lys-Gly-Ala-Pro-Glu-Arg。因此,在哇巴因敏感型和哇巴因不敏感型(钠,钾)-ATP酶中,5'-异硫氰酸荧光素反应位点的一级结构,因此大概还有ATP结合位点,似乎是完全保守的。