利用针对属于该酶反应中心一部分的合成肽段的抗体检测大鼠肾脏γ-谷氨酰转肽酶的构象变化。
Detection of conformational changes in rat kidney gamma-glutamyl transpeptidase by an antibody against a synthetic peptide belonging to part of the reactive centre of the enzyme.
作者信息
Glass G A, Bluvshtein E, Gur Y, Sfez S, Simovich V, Stark A A
机构信息
Department of Biochemistry, Tel-Aviv University, Ramat-Aviv, Israel.
出版信息
Biochem Mol Biol Int. 1994 Jun;33(3):505-13.
Polyclonal antibodies were produced against rat kidney gamma-glutamyl transpeptidase (GGT) and against a synthetic peptide corresponding to residues 512-534 in rat GGT. The anti-peptide antibody bound denatured GGT, acivicin-treated GGT and GGT absorbed to microtiter plates, but not GGT in solution, and did not inhibit GGT. The antibody against native GGT inhibited its activity in solution and did not bind efficiently adsorbed GGT in direct ELISA. It was active in direct and competition ELISA using kidney brush border membranes as the adsorbed antigen. The results indicate that these antibodies recognize conformational rather than sequence epitopes in GGT, and that marked changes occur in the conformation of GGT upon its absorption to plates or its reaction with acivicin.
制备了针对大鼠肾脏γ-谷氨酰转肽酶（GGT）以及针对与大鼠GGT中512 - 534位残基相对应的合成肽的多克隆抗体。抗肽抗体能结合变性的GGT、阿西维辛处理过的GGT以及吸附在微量滴定板上的GGT，但不能结合溶液中的GGT，且不抑制GGT活性。抗天然GGT抗体可抑制溶液中GGT的活性，在直接ELISA中不能有效结合吸附的GGT。在以肾刷状缘膜作为吸附抗原的直接ELISA和竞争ELISA中，该抗体具有活性。结果表明，这些抗体识别的是GGT中的构象表位而非序列表位，并且GGT在吸附到板上或与阿西维辛反应后其构象会发生显著变化。
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