Purification of a major sialoglycoprotein (SGP140) on P12/Ichikawa cells and its expression on differentiated HL-60 cells.
Purification of a major sialoglycoprotein (SGP140) on P12/Ichikawa cells and its expression on differentiated HL-60 cells.
复制标题
P12/Ichikawa 细胞上主要唾液酸糖蛋白 (SGP140) 的纯化及其在分化的 HL-60 细胞上的表达。
DOI：
10.4049/jimmunol.132.1.340
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发表时间：
1984
影响因子：
4.4
通讯作者：
Y. Nishikawa
中科院分区：
文献类型：
--
作者：
Y. Yamamoto;H. Mitsui;Y. Nishikawa
SGP 140糖蛋白是一种主要的细胞表面唾液酸糖蛋白，具有明显的分子量。用高碘酸盐-氚化硼氢化钠标记人T淋巴母细胞系P12/Ichikawa，然后用尿素-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和荧光照相法检测到140，000。然后从P12/Ichikawa细胞中纯化Sgp 140，研究其生化特性及其在不同细胞系中的分布。经0.2%TritonX-100溶解、DEAE-Sephacel柱层析、WGA-琼脂糖柱层析、Blue-Sepharose 6 MB柱层析和Sephadex G-150凝胶过滤进行纯化。然后制备抗SGP 140的抗血清，并对各种细胞系进行免疫沉淀和膜免疫荧光测定。在P12/Ichikawa、Raji、P3 HR-1、Daudi、Namalva、BALL-1、MOLT-4 B、TALL-1、NALL-1和K562细胞上检测到Sgp 140，但在HL-60细胞上未检测到。当HL-60细胞用二甲基亚砜，视黄酸，或12-O-十四烷酰基佛波醇-13-乙酸酯处理时，在细胞表面检测到SGP 140。我们讨论了Sgp 140可能是一种分化抗原的可能性。
SGP140 glycoprotein, a major cell surface sialoglycoprotein with an apparent m.w. of 140,000, was detected on the human T lymphoblastoid cell line P12/Ichikawa by labeling with periodate-tritiated sodium borohydride, followed by urea-sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Then SGP140 was purified from P12/Ichikawa cells for study of its biochemical character and its distribution in various cell lines. The purification was performed by 0.2% Triton X-100 solubilization from crude membranes, DEAE-Sephacel column chromatography, WGA-agarose column chromatography, Blue-Sepharose 6MB column chromatography, and Sephadex G-150 gel filtration. Antiserum raised against SGP140 was then prepared, and immunoprecipitation and membrane immunofluorescence assay were performed on various cell lines. SGP140 was detected on P12/Ichikawa, Raji, P3HR-1, Daudi, Namalva, BALL-1, MOLT-4B, TALL-1, NALL-1, and K562 cells, but was not detected on HL-60 cells. When HL-60 cells were treated with dimethyl sulfoxide, retinoic acid, or 12-O-tetradecanoylphorbol-13-acetate, SGP140 was detected on cell surfaces. We discuss the possibility that SGP140 may be a differentiation antigen.