Molecular characterization of two human autoantigens: unique cDNAs encoding 95- and 160-kD proteins of a putative family in the Golgi complex

Molecular characterization of two human autoantigens: unique cDNAs encoding 95- and 160-kD proteins of a putative family in the Golgi complex
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两种人类自身抗原的分子特征:编码高尔基复合体中推定家族的 95-和 160-kD 蛋白质的独特 cDNA

DOI:
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发表时间:
1993
影响因子:
15.3
通讯作者:
E. K. Chan
E. K. Chan
中科院分区:
医学1区
文献类型:
--
作者:
M. J. Fritzler;J. Hamel;R. Ochs;E. K. Chan

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使用来自具有针对高尔基复合体的自身抗体的患者的血清自身抗体来筛选来自 HepG2 lambda Zap cDNA 文库的克隆。纯化编码两种不同多肽的三个相关克隆,命名为 SY2、SY10 和 SY11,用于进一步分析。通过吸附到 lambda Zap 亲和纯化的抗体 - 克隆的重组蛋白和来自用纯化的重组蛋白免疫的 NZW 兔的抗体再现了高尔基体染色并结合了来自全细胞提取物的两种不同的蛋白,95 和 160 kD。 SY11蛋白暂命名为golgin-95,SY2/SY10蛋白暂命名为golgin-160。 SY2和SY11的cDNA克隆的推导的氨基酸序列代表568和620个氨基酸的58.7-和70-kD蛋白质。 SY2 和 SY11 cDNA 的体外翻译产物在 SDS-PAGE 中分别以 65 和 95 kD 迁移。体外翻译的蛋白通过人抗高尔基体血清或免疫兔血清进行免疫沉淀,但不能通过正常人血清或免疫前兔血清进行免疫沉淀。 cDNA的特征表明SY11是编码golgin-95的全长克隆,但SY2和SY10一起编码golgin-160的部分序列。对SY11重组蛋白的分析鉴定出跨越位置419-455的亮氨酸拉链、跨越位置322-333的谷氨酸富集区和跨越位置67-73的富含脯氨酸的区。对 SwissProt 数据库的检索表明 SY11 与人休息蛋白、驱动蛋白重链和肌球蛋白重链的序列相似。 SY2 与肌球蛋白的重链、来自酵母的 USO1 转运蛋白和 150 kD 细胞质动力蛋白相关多肽具有序列相似性。序列分析表明,golgin-95 和 golgin-160 具有 43% 的序列相似性,因此可能是功能相关的蛋白质。
Serum autoantibodies from a patient with autoantibodies directed against the Golgi complex were used to screen clones from a HepG2 lambda Zap cDNA library. Three related clones, designated SY2, SY10, and SY11, encoding two distinct polypeptides were purified for further analysis. Antibodies affinity purified by adsorption to the lambda Zap- cloned recombinant proteins and antibodies from NZW rabbits immunized with purified recombinant proteins reproduced Golgi staining and bound two different proteins, 95 and 160 kD, from whole cell extracts. The SY11 protein was provisionally named golgin-95 and the SY2/SY10 protein was named golgin-160. The deduced amino acid sequence of the cDNA clone of SY2 and SY11 represented 58.7- and 70-kD proteins of 568 and 620 amino acids. The in vitro translation products of SY2 and SY11 cDNAs migrated in SDS-PAGE at 65 and 95 kD, respectively. The in vitro translated proteins were immunoprecipitated by human anti-Golgi serum or immune rabbit serum, but not by normal human serum or preimmune rabbit serum. Features of the cDNA suggested that SY11 was a full- length clone encoding golgin-95 but SY2 and SY10 together encoded a partial sequence of golgin-160. Analysis of the SY11 recombinant protein identified a leucine zipper spanning positions 419-455, a glutamic acid-rich tract spanning positions 322-333, and a proline-rich tract spanning positions 67-73. A search of the SwissProt data bank indicated sequence similarity of SY11 to human restin, the heavy chain of kinesin, and the heavy chain of myosin. SY2 shared sequence similarity with the heavy chain of myosin, the USO1 transport protein from yeast, and the 150-kD cytoplasmic dynein-associated polypeptide. Sequence analysis demonstrated that golgin-95 and golgin-160 share 43% sequence similarity and, therefore, may be functionally related proteins.
参与 Friend 鼠白血病病毒包膜糖蛋白寡聚化的分子结构域。
DOI: 10.1016/0042-6822(91)90542-j
发表时间: 1991
期刊: Virology
影响因子: 3.7
作者:
Tucker,SP;Srinivas,RV;Compans,RW
通讯作者: Compans,RW
DOI: 10.1172/jci115003
发表时间: 1991-01-01
影响因子: 15.9
作者:
CHAN, EKL;HAMEL, JC;TAN, EM
通讯作者: TAN, EM
DOI: 10.1146/annurev.cb.02.110186.002005
发表时间: 1986
期刊: Annual review of cell biology
影响因子: --
作者:
S. Singer;A. Kupfer
通讯作者: S. Singer;A. Kupfer
Ku 抗原 86 kDa 亚基的 cDNA 衍生氨基酸序列。
DOI: --
发表时间: 1989
期刊: The Journal of biological chemistry
影响因子: --
作者:
Yaneva,M;Wen,J;Ayala,A;Cook,R
通讯作者: Cook,R
DOI: 10.1021/bi00409a043
发表时间: 1988-05-03
期刊: BIOCHEMISTRY
影响因子: 2.9
作者:
BLOOM, GS;WAGNER, MC;BRADY, ST
通讯作者: BRADY, ST