Molecular Characterization of trans-Golgi p230

Molecular Characterization of trans-Golgi p230
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反式高尔基体 p230 的分子表征

DOI:
--
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发表时间:
1996
影响因子:
4.8
通讯作者:
B. Toh
B. Toh
中科院分区:
生物学2区
文献类型:
--
作者:
R. Erlich;P. Gleeson;P. Campbell;E. Dietzsch;B. Toh

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使用来自干燥综合征患者的自身抗体,我们先前已经鉴定了与反式高尔基体的胞质面相关的230-kDa外周膜蛋白(Kooy,J.,托,B。H、佩蒂特,J.M.,埃利希河和Gleeson,P. A.(1992)J.Biol.Chem.267,20255-20263)。本文报道了人p230基因的克隆、序列分析及其在染色体6p 12 -22上的定位。使用自身抗体从HeLa细胞cDNA文库中分离的部分cDNA克隆用于获得额外的cDNA,其共同跨越7695个碱基对(bp)。p230 mRNA为7.7个酶。检测到两种p230的选择性剪接mRNA。这些差异在于3′序列中的21-和63-bp插入,导致羧基末端氨基酸序列的差异。预测的261-kDa蛋白是高度亲水性的,与许多含有卷曲螺旋结构域的蛋白质具有17-20%的同源性。除了两个富含脯氨酸的区域(氨基酸1-117和239-270)外,p230还含有非常高频率的七肽重复序列,这是形成二聚体卷曲螺旋结构的α螺旋的特征。p230还包括序列ESLALEEEL(氨基酸538-546),其是在存在于神经内分泌细胞的分泌颗粒中的酸性蛋白质的粒蛋白家族中发现的基序。这是第一次报告的细胞溶质高尔基体蛋白含有颗粒蛋白基序。p230的结构特征表明,它可能在从trans-Golgi的囊泡运输中发挥作用。
Using autoantibodies from a Sjögren's syndrome patient, we have previously identified a 230-kDa peripheral membrane protein associated with the cytosolic face of the trans-Golgi (Kooy, J., Toh, B. H., Pettitt, J. M., Erlich, R. and Gleeson, P. A.(1992) J. Biol. Chem. 267, 20255-20263). Here we report the molecular cloning and sequence analysis of human p230 and the localization of its gene to chromosome 6p12-22. Partial cDNA clones, isolated from a HeLa cell cDNA library using autoantibodies, were used to obtain additional cDNAs, which together span 7695 base pairs (bp). The p230 mRNA is 7.7 kilobases. Two alternatively spliced mRNAs for p230 were detected. These differed by 21- and 63-bp insertions in the 3′-sequence, resulting in differences in amino acid sequence at the carboxyl terminus. The predicted 261-kDa protein is highly hydrophilic with 17-20% homology with many proteins containing coiled-coil domains. Apart from two proline-rich regions (amino acids 1-117 and 239-270), p230 contains a very high frequency of heptad repeats, characteristic of α-helices that form dimeric coiled-coil structures. p230 also includes the sequence ESLALEELEL (amino acids 538-546), a motif found in the granin family of acidic proteins present in secretory granules of neuroendocrine cells. This is the first report of a cytosolic Golgi protein containing a granin motif. The structural characteristics of p230 indicate that it may play a role in vesicular transport from the trans-Golgi.
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