RAT-BRAIN CANNABINOID RECEPTORS ARE N-LINKED GLYCOSYLATED PROTEINS
RAT-BRAIN CANNABINOID RECEPTORS ARE N-LINKED GLYCOSYLATED PROTEINS
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DOI：
10.1016/0024-3205(95)00179-a
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发表时间：
1995-05-05
期刊：
影响因子：
6.1
通讯作者：
HOWLETT, AC
中科院分区：
文献类型：
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作者：
SONG, C;HOWLETT, AC
为了研究CB1受体的n链糖基化特性，用外糖苷酶和内糖苷酶处理大鼠脑膜。为了可视化CB1受体，针对n端14个氨基酸提出了一种抗肽抗体，并通过Western blotting特异性检测该蛋白。我们发现成熟CB1受体的表观分子量为64 kDa。用内糖苷酶F处理膜将64 kDa带转移到59 kDa和53 kDa带。后者与去糖基化CB1受体的计算分子量一致。用内糖苷酶H和α -甘露糖苷酶处理膜后，64 kDa带部分移位到53 kDa带，表明部分低聚糖为高甘露糖型。这些数据证实了脑内CB1受体是具有异质碳水化合物组成的n -连接糖蛋白。在CB1受体n端上的三个潜在的n链糖基化位点中，只有两个位点实际上被糖基化了。
To study the N-linked glycosylation properties of the CB1 receptor, rat brain membranes were treated with exo- and endoglycosidases. For visualizing CB1 receptors, an antipeptide antibody was raised against the N-terminal 14 amino acids and used to specifically detect the protein by Western blotting., We found that the apparent molecular weight of mature CB1 receptors was 64 kDa. Treatment of membranes with endoglycosidase F shifted the 64 kDa band to the 59 kDa and 53 kDa bands. The latter is consistent with the calculated molecular weight of deglycosylated CB1 receptors. Treatment of membranes with endoglycosidase H and alpha-mannosidase partially shifted the 64 kDa band to 53 kDa band, indicating a portion of the oligosaccharides was of the high mannose type. These data confirmed that the CB1 receptors in brain are N-linked glycoproteins with heterogeneous carbohydrate composition. Among three potential N-linked glycosylation sites on the N-terminus of the CB1 receptor, only two sites are actually glycosylated.