Identification and purification of a Mr 75,000 cell surface human melanoma-associated antigen.
Identification and purification of a Mr 75,000 cell surface human melanoma-associated antigen.
复制标题
Mr 75,000 细胞表面人类黑色素瘤相关抗原的鉴定和纯化。
作者：
J. Heaney;J. Bystryn
通过乳过氧化物酶方法对培养的人黑色素瘤细胞的表面大分子进行放射性碘标记。收集、浓缩流入培养基中的大分子，并在 Sepharose 6B 和扁豆凝集素-Sepharose 上分级。通过三氯乙酸沉淀来测定与大分子相关的放射性，并通过异种抗黑色素瘤血清的特异性免疫沉淀来测定与黑色素瘤相关抗原（MAA）相关的放射性。在纯化的最后一步之后，MAA 活性最强的级分中大约三分之二的酸不溶性放射性与该抗原相关。该级分中的 MAA 浓度比细胞中的 MAA 浓度高约 200 倍，并且通过在十二烷基硫酸钠中进行聚丙烯酰胺凝胶电泳，其包含单个标记的蛋白质条带。 MAA是一种由单条多肽链组成的糖蛋白，分子量约为75,000。这种抗原是几种但不是所有黑色素瘤所共有的。在 16 个其他不相关的同种异体或异种正常和恶性细胞中未检测到它，或者其浓度大大降低。它在免疫学上与血清白蛋白或纤连蛋白无关。这些观察结果表明，我们高度纯化了人类黑色素瘤细胞的 Mr 75,000 细胞表面成分，它似乎是 MAA。
Surface macromolecules on cultured human melanoma cells wee radioiodinated by a lactoperoxidase method. Macromolecules shed into culture medium were collected, concentrated, and fractionated on Sepharose 6B and lentil lectin-Sepharose. Radioactivity associated with macromolecules was assayed by precipitation with trichloracetic acid, and that associated with melanoma-associated antigens (MAA) was assayed by specific immunoprecipitation with xenogeneic anti-melanoma serum. Following the last step in purification, approximately two-thirds of the acid-insoluble radioactivity in the most MAA-active fraction was associated with this antigen. MAA concentration in this fraction was approximately 200-fold greater than that in cells, and it contained a single labeled protein band by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The MAA was a glycoprotein consisting of a single polypeptide chain with a molecular weight of approximately 75,000. This antigen was common to several but not to all melanomas. It was not detected, or was present in much decreased concentration, in 16 other unrelated allogeneic or xenogeneic normal and malignant cells. It was immunologically unrelated to serum albumin or fibronectin. These observations indicate that we have highly purified a Mr 75,000 cell surface component of human melanoma cell which appears to be a MAA.
影响因子：
11.2
作者：
M. Herlyn;W. Clark;M. Mastrangelo;D. Guerry;D. Elder;D. Larossa;R. Hamilton;E. Bondi;R. Tuthill;Z. Steplewski;H. Koprowski
通讯作者：
M. Herlyn;W. Clark;M. Mastrangelo;D. Guerry;D. Elder;D. Larossa;R. Hamilton;E. Bondi;R. Tuthill;Z. Steplewski;H. Koprowski
人黑色素瘤细胞表面的生化特征：用单克隆抗体解剖。
DOI：
10.1073/pnas.77.12.7287
发表时间：
1980
影响因子：
11.1
作者：
Mitchell,KF;Fuhrer,JP;Steplewski,Z;Koprowski,H
通讯作者：
Koprowski,H