RELATIONSHIP BETWEEN GLYCOCALICIN AND GLYCOPROTEIN IB OF HUMAN-PLATELETS
RELATIONSHIP BETWEEN GLYCOCALICIN AND GLYCOPROTEIN IB OF HUMAN-PLATELETS
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DOI：
10.1073/pnas.78.5.2712
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
LUSCHER, EF
中科院分区：
文献类型：
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作者：
CLEMETSON, KJ;NAIM, HY;LUSCHER, EF
用花生凝集素亲和层析法，将神经氨酸酶处理的人血小板超声处理后，分别从膜和上清液中分离得到去唾液酸糖蛋白Ib和去唾液糖钙蛋白。去唾液酸糖蛋白Ib在未还原时的表观相对分子质量为16万，还原时的表观相对分子质量为15万；去唾液酸糖钙蛋白在十二烷基硫酸钠/聚丙烯酰胺凝胶上的表观相对分子质量为15万，包括还原和未还原。两种制剂都只含有微量的杂质。用凝胶电泳法分离未还原和还原状态的去唾液酸糖蛋白Ib和去唾液酸糖钙蛋白，凝胶切片进行碘标记，胰酶消化，用二维高压电泳法和薄层层析结合放射自显影进行分析。胰酶肽谱显示糖钙素和糖蛋白Ib有很大的相似性，后者(未还原和还原)都含有额外的多肽，支持糖钙素来自糖蛋白Ib的观点。未还原的糖蛋白Ib与由于二硫键连接的β-β而还原的糖蛋白Ib相比，含有额外的多肽。组件。未还原糖钙蛋白和还原糖钙蛋白也略有不同，表明至少存在一个分子内二硫键。
Asialoglycoprotein Ib and asialoglycocalicin were isolated from the membranes and from the supernatant, respectively, after sonication of neuraminidase-treated [human] platelets, by lectin affinity chromatography on peanut agglutinin. The isolated asialoglycoprotein Ib had an apparent MW of 160,000 when not reduced and 150,000 when reduced; the asialoglycocalicin had an apparent MW of 150,000, both reduced and unreduced, on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Both preparations contained only trace amounts of impurities. The asialoglycoprotein Ib and asialoglycocalicin in both the unreduced and reduced states were separated by gel electrophoresis, redioiodinated in gel slices and digested with trypsin, and the digests were analyzed by 2-dimensional high-voltage electrophoresis and TLC followed by autoradiography. The tryptic peptide maps showed great similarities between glycocalicin and glycoprotein Ib, with the latter (both unreduced and reduced) containing additional peptides, supporting the idea that glycocalicin is derived from glycoprotein Ib. The unreduced glycoprotein Ib contained additional peptides compared to the reduced due to the disulfide-bond-linked .beta. component. There were also slight differences between unreduced and reduced glycocalicin, indicating that at least 1 intramolecular disulfide bond is present.