Tissue distribution, structural characterization, and biosynthesis of Mac-3, a macrophage surface glycoprotein exhibiting molecular weight heterogeneity.
Tissue distribution, structural characterization, and biosynthesis of Mac-3, a macrophage surface glycoprotein exhibiting molecular weight heterogeneity.
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DOI：
10.1016/s0021-9258(18)33302-7
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发表时间：
1983-01
期刊：
影响因子：
--
通讯作者：
M. Ho;T. Springer
中科院分区：
文献类型：
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作者：
M. Ho;T. Springer
Mac-3是由大鼠抗小鼠单克隆抗体（MAb）M3/84定义的小鼠巨噬细胞分化抗原。Mac-3的结构，生物合成，定量表面表达和分布进行了研究，通过放射性标记和分离与MAb-Sepharose，饱和结合，吸收，免疫荧光流式细胞术。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，Mac-3以平均Mr = 110，000的扩散带迁移。用125 I标记完整细胞和MAb的可接近性表明它至少部分存在于细胞表面。125 I-MAb饱和标记显示巯基乙酸盐培养基诱导的腹腔巨噬细胞上有4.2 × 10（4）个细胞表面位点。[35 S]蛋氨酸和[3 H]葡糖胺通过纯化的巨噬细胞掺入Mac-3中，表明它是由这些细胞合成的糖蛋白。吸收显示Mac-3在巨噬细胞中最强，在肺、肝、骨髓和脾中含量较低，在胸腺、淋巴结、脑和心脏中检测不到。免疫荧光流式细胞术显示巯基乙酸盐诱导的巨噬细胞表面表达，但骨髓、脾脏、淋巴结或胸腺细胞悬液不表达。类似数量的Mac-3从驻留的巨噬细胞或由无菌炎症剂、细胞内寄生虫或免疫调节剂引起的巨噬细胞中免疫沉淀，但Mac-3的平均Mr从92，000到110，000不等。Mac-3由Mr = 74，000和79，000的前体合成，在不同的巨噬细胞中相同。在15至30分钟内，成熟分子被加工成成熟分子，该成熟分子在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中迁移为更弥散的条带，并且在不同试剂引起的巨噬细胞之间的Mr不同，并且在同一类型巨噬细胞的不同制剂之间的Mr不同程度上不同。
Mac-3 is a mouse macrophage differentiation antigen defined by a rat anti-mouse monoclonal antibody (MAb),M3/84. The structure, biosynthesis, quantitative surface expression, and distribution of Mac-3 have been studied by radiolabeling and isolation with MAb-Sepharose, saturation binding, absorption, and immunofluorescence flow cytometry. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Mac-3 migrates as a diffuse band with average Mr = 110,000. Labeling of intact cells with 125I and accessibility to MAb show it is present at least in part on the cell surface. Saturation labeling with 125I-MAb shows 4.2 X 10(4) cell surface sites on thioglycollate medium-elicited peritoneal macrophages. [35S]Methionine and [3H]glucosamine incorporation into Mac-3 by purified macrophages show it is a glycoprotein synthesized by these cells. Absorption shows Mac-3 is strongest in macrophages, present in lower quantities in lung, liver, bone marrow, and spleen, and undetectable in thymus, lymph node, brain, and heart. Immunofluorescent flow cytometry shows surface expression on thioglycollate-elicited macrophages but not bone marrow, spleen, lymph node, or thymus cell suspensions. Similar amounts of Mac-3 are immunoprecipitated from resident macrophages or macrophages elicited by sterile inflammatory agents, intracellular parasites, or immunomodulators, but the average Mr of Mac-3 varies from 92,000 to 110,000. Mac-3 is synthesized from precursor(s) of Mr = 74,000 and 79,000, identical in the different macrophages. Processing into the mature molecule, which migrates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a more diffuse band and varies in Mr among macrophage elicited by different agents and to a lesser degree between different preparations of the same type of macrophage, occurs in 15 to 30 min.