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编码gp138的两个基因的分子克隆与特性分析,gp138是一种参与盘基网柄菌有性细胞融合的细胞表面糖蛋白。

Molecular cloning and characterization of two genes encoding gp138, a cell surface glycoprotein involved in the sexual cell fusion of Dictyostelium discoideum.

作者信息

Fang H, Higa M, Suzuki K, Aiba K, Urushihara H, Yanagisawa K

机构信息

Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.

出版信息

Dev Biol. 1993 Mar;156(1):201-8. doi: 10.1006/dbio.1993.1070.

Abstract

The sexual development of cellular slime molds is initiated by acquisition of sexual fusion competence of myxamoeboid cells. During the acquisition of fusion competence in NC4 and HM1 strain cells of Dictyostelium discoideum, opposite in mating types, a glycoprotein gp138, relevant to sexual cell fusion, is expressed on the cell surfaces of both the strains. To investigate the mechanisms controlling cell fusion, gp138 was purified and its N-terminal amino acid sequence was determined. An oligonucleotide with sequence predicted from the amino acid sequence was synthesized to isolate the gene encoding gp138. Two genes, referred to as GP138A and GP138B, were then cloned. They contain the region which encodes the N-terminal amino acid sequence of the gp138 protein and their structure is very similar on the whole. The C-terminal portion of the predictive polypeptides produced by the genes is highly hydrophobic and proline rich. It has homology with a portion of gp80, a glycoprotein for cell-cell adhesion, and PsA, a glycoprotein specific for prespore cells, reported previously in D. discoideum. The expression of GP138A is greater than that of GP138B. The mRNA of GP138A is expressed at the time of acquisition of fusion competence of cells during cultivation but the mRNA of GP138B is not.

摘要

细胞黏菌的有性发育是由黏变形体细胞获得性融合能力引发的。在盘基网柄菌的NC4和HM1菌株细胞(交配型相反)获得融合能力的过程中,与性细胞融合相关的糖蛋白gp138在这两种菌株的细胞表面表达。为了研究控制细胞融合的机制,对gp138进行了纯化并测定了其N端氨基酸序列。根据氨基酸序列预测合成了一段寡核苷酸,用于分离编码gp138的基因。随后克隆了两个基因,分别称为GP138A和GP138B。它们包含编码gp138蛋白N端氨基酸序列的区域,整体结构非常相似。这些基因产生的预测多肽的C端部分高度疏水且富含脯氨酸。它与先前在盘基网柄菌中报道的细胞间黏附糖蛋白gp80以及前孢子细胞特异性糖蛋白PsA的一部分具有同源性。GP138A的表达量大于GP138B。GP138A的mRNA在培养过程中细胞获得融合能力时表达,而GP138B的mRNA则不表达。

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