Glycoproteins of the lysosomal membrane.
Glycoproteins of the lysosomal membrane.
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溶酶体膜的糖蛋白。
DOI：
10.1083/jcb.100.6.1839
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发表时间：
1985-06
影响因子：
7.8
通讯作者：
MELLMAN, I
中科院分区：
文献类型：
--
作者：
LEWIS, V;GREEN, SA;MARSH, M;VIHKO, P;HELENIUS, A;MELLMAN, I
三个糖蛋白抗原（120，100，和80 kD）检测单和/或多克隆抗体免疫与高度纯化的大鼠肝溶酶体膜。根据Triton X-114的结合，判断所有抗原均为膜整合蛋白。通过对正常大鼠肾细胞的免疫荧光，小鼠单克隆抗体的120-kD抗原共染色的多克隆兔抗体，检测100-和80-kD抗原，以及与抗体酸性磷酸酶，表明这些抗原优先定位于溶酶体。少数120 kD阳性结构被发现是阴性的酸性磷酸酶，这表明抗原不集中在细胞器，如内体，缺乏酸性磷酸酶。免疫过氧化物酶细胞化学也表明，在高尔基体池，包被囊泡，或质膜上的反应性很小。用内切-β-N-乙酰氨基葡萄糖苷酶H（Endo H）和内切-β-N-乙酰氨基葡萄糖苷酶F（Endo F）消化证明，每种抗原含有多个N-连接的寡糖链，其中大多数是复合物（Endo H-抗性）类型。120-kD蛋白是非常严重的糖基化，具有至少18个N-连接的链。它也富含唾液酸，因为神经氨酸酶消化增加了120 kD蛋白质的pI从小于4到大于8。两者合计，这些结果强烈表明，溶酶体膜的糖蛋白组分在粗面内质网中合成，并在高尔基体中最终糖基化，然后递送到溶酶体。我们暂时将这些抗原命名为溶酶体膜糖蛋白lgp 120、lgp 100、lgp 80。
Three glycoprotein antigens (120, 100, and 80 kD) were detected by mono- and/or polyclonal antibodies generated by immunization with highly purified rat liver lysosomal membranes. All of the antigens were judged to be integral membrane proteins based on the binding of Triton X-114. By immunofluorescence on normal rat kidney cells, a mouse monoclonal antibody to the 120-kD antigen co-stained with a polyclonal rabbit antibody that detected the 100- and 80-kD antigens as well as with antibodies to acid phosphatase, indicating that these antigens are preferentially localized in lysosomes. Few 120-kD-positive structures were found to be negative for acid phosphatase, suggesting that the antigen was not concentrated in organelles such as endosomes, which lack acid phosphatase. Immunoperoxidase cytochemistry also showed little reactivity in Golgi cisternae, coated vesicles, or on the plasma membrane. Digestion with endo-beta-N-acetylglucosaminidase H (Endo H) and endo-beta-N-acetylglucosaminidase F (Endo F) demonstrated that each of the antigens contained multiple N-linked oligosaccharide chains, most of which were of the complex (Endo H-resistant) type. The 120-kD protein was very heavily glycosylated, having at least 18 N-linked chains. It was also rich in sialic acid, since neuraminidase digestion increased the pI of the 120-kD protein from less than 4 to greater than 8. Taken together, these results strongly suggest that the glycoprotein components of the lysosomal membrane are synthesized in the rough endoplasmic reticulum and terminally glycosylated in the Golgi before delivery to lysosomes. We have provisionally designated these antigens lysosomal membrane glycoproteins lgp120, lgp100, lgp80.
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DOI：
10.1073/pnas.79.15.4540
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ELDER, JH;ALEXANDER, S
通讯作者：
ALEXANDER, S
影响因子：
7.8
作者：
HUBBARD, AL;COHN, ZA
通讯作者：
COHN, ZA
影响因子：
7.8
作者：
MEHRABIAN, M;BAME, KJ;ROME, LH
通讯作者：
ROME, LH
DOI：
10.1073/pnas.80.3.775
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
GABEL, CA;GOLDBERG, DE;KORNFELD, S
通讯作者：
KORNFELD, S
影响因子：
2.9
作者：
CHAMBERLAIN, JP
通讯作者：
CHAMBERLAIN, JP