Characterization of a cis-Golgi matrix protein, GM130.
Characterization of a cis-Golgi matrix protein, GM130.
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DOI：
10.1083/jcb.131.6.1715
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发表时间：
1995-12
期刊：
影响因子：
--
通讯作者：
Warren G
中科院分区：
文献类型：
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作者：
Nakamura N;Rabouille C;Watson R;Nilsson T;Hui N;Slusarewicz P;Kreis TE;Warren G
抗血清提出的洗涤剂和盐的抗基质部分从大鼠肝脏高尔基体堆栈被用来筛选大鼠肝脏cDNA的表达文库。获得编码130 kD蛋白质（称为GM 130）的全长克隆，其COOH末端结构域与高尔基体人自身抗原golgin-95高度同源（Fritzler等人，1993年）。生物化学数据表明，GM 130是一种外周细胞质蛋白，与高尔基体膜紧密结合，是一个更大的寡聚复合物的一部分。从蛋白质序列的预测表明，GM 130是一个扩展的棒状蛋白与卷曲螺旋结构域。免疫荧光显微镜显示部分重叠的中间和反式高尔基体标记，但几乎完全重叠的顺式高尔基体网络（CGN）标记，突触融合蛋白5。免疫电子显微镜证实了这一位置，显示大部分的GM 130位于CGN和一个或两个池的顺侧的高尔基体堆栈。GM 130在布雷菲德菌素A存在下不重新分布到ER，但保持其与突触融合蛋白5的重叠和与ER-高尔基体中间室标记物p53的部分重叠.总之，这些结果表明，GM 130是一个顺式高尔基体基质的一部分，并在维持顺式高尔基体结构的作用。
Antisera raised to a detergent- and salt-resistant matrix fraction from rat liver Golgi stacks were used to screen an expression library from rat liver cDNA. A full-length clone was obtained encoding a protein of 130 kD (termed GM130), the COOH-terminal domain of which was highly homologous to a Golgi human auto-antigen, golgin-95 (Fritzler et al., 1993). Biochemical data showed that GM130 is a peripheral cytoplasmic protein that is tightly bound to Golgi membranes and part of a larger oligomeric complex. Predictions from the protein sequence suggest that GM130 is an extended rod-like protein with coiled-coil domains. Immunofluorescence microscopy showed partial overlap with medial- and trans-Golgi markers but almost complete overlap with the cis-Golgi network (CGN) marker, syntaxin5. Immunoelectron microscopy confirmed this location showing that most of the GM130 was located in the CGN and in one or two cisternae on the cis-side of the Golgi stack. GM130 was not re-distributed to the ER in the presence of brefeldin A but maintained its overlap with syntaxin5 and a partial overlap with the ER- Golgi intermediate compartment marker, p53. Together these results suggest that GM130 is part of a cis-Golgi matrix and has a role in maintaining cis-Golgi structure.