Stricker R, Kalbacher H, Lottspeich F, Reiser G
Institut für Neurobiochemie der Otto-von-Guericke Universität Magdeburg, Germany.
FEBS Lett. 1995 Aug 21;370(3):236-40. doi: 10.1016/0014-5793(95)00822-q.
From a high-affinity Ins-P4 (inositol 1,3,4,5-P4) receptor purified from pig cerebellum, digested with the protease Lys C peptide sequences were obtained. Synthetic peptide-3 (19 amino acid residues) was used to generate an antiserum. Reaction of the affinity-purified antibodies with the purified pig receptor protein in ELISA or Western blot was completely inhibited by peptide-3. In cerebellar membranes, the antibodies clearly recognized the 42 kDa Ins-P4 receptor protein and two additional proteins (25 kDa, 37 kDa) which still have to be identified. The anti-peptide antibodies could selectively immunoprecipitate the Ins-P4 receptor protein. The antiserum was used (i) to demonstrate that in brain from different species (human, pig, beef, rat, mouse and sheep) a similar 42 kDa Ins-P4 receptor protein is contained, and (ii) to obtain indications for the existence of a related soluble form of the 42 kDa Ins-P4 receptor besides the membrane-associated receptor.
从小猪小脑纯化的高亲和力肌醇-1,3,4,5-四磷酸（Ins-P4）受体中，经蛋白酶Lys C消化后获得了肽序列。合成肽-3（19个氨基酸残基）用于制备抗血清。在ELISA或蛋白质印迹中，亲和纯化的抗体与纯化的猪受体蛋白的反应被肽-3完全抑制。在小脑膜中，抗体能清晰识别42 kDa的Ins-P4受体蛋白以及另外两种有待鉴定的蛋白（25 kDa、37 kDa）。抗肽抗体能够选择性地免疫沉淀Ins-P4受体蛋白。该抗血清被用于：（i）证明在不同物种（人类、猪、牛、大鼠、小鼠和绵羊）的大脑中都含有类似的42 kDa Ins-P4受体蛋白；（ii）获取除膜相关受体外，42 kDa Ins-P4受体相关可溶性形式存在的证据。
