Identification of two lysosomal membrane glycoproteins.
Identification of two lysosomal membrane glycoproteins.
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DOI：
10.1083/jcb.101.1.85
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发表时间：
1985-07
影响因子：
7.8
通讯作者：
AUGUST, JT
中科院分区：
文献类型：
--
作者：
CHEN, JW;MURPHY, TL;WILLINGHAM, MC;PASTAN, I;AUGUST, JT
特异性结合溶酶体膜的单克隆抗体已鉴定出两种小鼠溶酶体相关膜蛋白，即 105,000-115,000 D 的 LAMP-1 和 100,000-110,000 D 的 LAMP-2。两种糖蛋白均被区分为可被去垢剂溶液溶解但不被各种离液剂溶解的完整膜成分。通过间接免疫荧光染色、抗原与吖啶橙摄取位点的共定位以及免疫电子显微镜证实了溶酶体定位。抗体结合主要位于溶酶体的限制膜上，与长时间孵育期间在管腔中积累的胶体金标记的 α-2-巨球蛋白明显分离。 LAMP-1 和 LAMP-2 似乎也以低浓度存在于高尔基体反式元件上，但在以新内吞的 α-2-巨球蛋白的存在为标志的受体体或其他细胞结构中未检测到。通过二维凝胶分析、 125 I 胰蛋白酶肽图谱和 125 I 标记细胞提取物的连续免疫沉淀，将 LAMP-1 和 LAMP-2 区分为不同的分子。两种糖蛋白都是作为大约 90,000 D 的前体蛋白合成的，并且在不同细胞系中表现出明显的表观分子量表达的异质性。抗体吸附和胰蛋白酶肽图谱表明，LAMP-2 与巨噬细胞抗原 MAC-3 密切相关或相同。据推测，这些糖蛋白作为溶酶体膜的主要蛋白质成分，在溶酶体结构和功能中具有重要作用。
Two murine lysosome-associated membrane proteins, LAMP-1 of 105,000- 115,000 D and LAMP-2 of 100,000-110,000 D, have been identified by monoclonal antibodies that bind specifically to lysosomal membranes. Both glycoproteins were distinguished as integral membrane components solubilized by detergent solutions but not by various chaotropic agents. The lysosome localization was demonstrated by indirect immunofluorescent staining, co-localization of the antigen to sites of acridine orange uptake, and immunoelectron microscopy. Antibody binding was predominantly located at the limiting lysosomal membrane, distinctly separated from colloidal gold-labeled alpha-2-macroglobulin accumulated in the lumen during prolonged incubation. LAMP-1 and LAMP-2 also appeared to be present in low concentrations on Golgi trans- elements but were not detected in receptosomes marked by the presence of newly endocytosed alpha-2-macroglobulin, or in other cellular structures. LAMP-1 and LAMP-2 were distinguished as different molecules by two-dimensional gel analysis, 125I-tryptic peptide mapping, and sequential immunoprecipitations of 125I-labeled cell extracts. Both glycoproteins were synthesized as a precursor protein of approximately 90,000 D, and showed a marked heterogeneity of apparent molecular weight expression in different cell lines. LAMP-2 was closely related or identical to the macrophage antigen, MAC-3, as indicated by antibody adsorption and tryptic peptide mapping. It is postulated that these glycoproteins, as major protein constituents of the lysosomal membrane, have important roles in lysosomal structure and function.
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DOI：
10.1073/pnas.77.2.990
发表时间：
1980-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
JOHNSON, LV;WALSH, ML;CHEN, LB
通讯作者：
CHEN, LB
影响因子：
5.4
作者：
SCOLNICK, EM;PARKS, WP
通讯作者：
PARKS, WP
DOI：
10.1111/j.1432-1033.1974.tb03599.x
发表时间：
1974-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BONNER, WM;LASKEY, RA
通讯作者：
LASKEY, RA
影响因子：
4.1
作者：
BURNSIDE, J;SCHNEIDER, DL
通讯作者：
SCHNEIDER, DL
影响因子：
64.8
作者：
LAEMMLI, UK
通讯作者：
LAEMMLI, UK