Kitlar T, Morrison A I, Kinne R, Deutscher J
Max-Planck-Institut für Systemphysiologie, Dortmund, FRG.
FEBS Lett. 1988 Jul 4;234(1):115-9. doi: 10.1016/0014-5793(88)81315-2.
Phlorizin, a potent inhibitor of the Na+/D-glucose cotransporter, was derivatised to 3-aminophlorizin and subsequently coupled to Affi-Gel 15. Affinity chromatography of pig kidney brush border membranes solubilised in Triton X-100 allowed the purification of a 60 kDa protein on this resin. We consider this protein to be the Na+/D-glucose cotransporter, or part of it, for the following reasons: (i) binding of this protein to Affi-Gel 15 specifically requires phlorizin covalently attached to the resin and is lowered when phlorizin is replaced by phloretin; (ii) binding of the 60 kDa protein to a phlorizin affinity column requires the presence of Na+; (iii) polyclonal as well as monoclonal antibodies against the 60 kDa protein inhibit binding of phlorizin to brush border membranes from rabbit and pig kidney.
根皮苷是一种有效的钠/葡萄糖共转运蛋白抑制剂，它被衍生化为3-氨基根皮苷，随后与Affi-Gel 15偶联。用Triton X-100溶解猪肾刷状缘膜后进行亲和层析，可在该树脂上纯化出一种60 kDa的蛋白质。我们认为该蛋白质是钠/葡萄糖共转运蛋白，或其一部分，原因如下：(i) 该蛋白质与Affi-Gel 15的结合特别需要共价连接在树脂上的根皮苷，当根皮苷被根皮素取代时结合会降低；(ii) 60 kDa蛋白质与根皮苷亲和柱的结合需要有Na+存在；(iii) 针对60 kDa蛋白质的多克隆抗体和单克隆抗体均能抑制根皮苷与兔肾和猪肾刷状缘膜的结合。
