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秀丽隐杆线虫的两种寡肽转运蛋白:分子克隆与功能表达。

Two oligopeptide transporters from Caenorhabditis elegans: molecular cloning and functional expression.

作者信息

Fei Y J, Fujita T, Lapp D F, Ganapathy V, Leibach F H

机构信息

Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, GA 30912, USA.

出版信息

Biochem J. 1998 Jun 1;332 ( Pt 2)(Pt 2):565-72. doi: 10.1042/bj3320565.

Abstract

Two novel oligopeptide transporter cDNA clones, CPTA and CPTB, were identified by screening a Caenorhabditis elegans cDNA library using homology hybridization. The transporter proteins deduced from the cDNAs possess multiple transmembrane domains and reveal a moderate similarity to their mammalian counterparts in amino acid sequences. CPTA and CPTB, when expressed in Xenopus laevis oocytes and studied by both radiotracer flux and microelectrode voltage-clamp protocol, displayed a saturable electrogenic transport activity driven by a proton gradient with an overlapping broad spectrum of substrate specificity. Both transporters recognize di-, tri- and tetra-peptides including phenylalanylmethionylarginylphenylalaninamide (FMRFamide) and N-acetylaspartylglutamate, members of a large neuropeptide family commonly found throughout the animal kingdom. Kinetic analysis, however, revealed that CPTA and CPTB differed in their affinity for the peptide substrates, the former being a high-affinity type and the latter a low-affinity type. CPTA and CPTB are encoded by two distinct genes localized on separate chromosomes and are expressed during the whole life span of the organism.

摘要

通过同源杂交筛选秀丽隐杆线虫cDNA文库,鉴定出两个新的寡肽转运体cDNA克隆,即CPTA和CPTB。从这些cDNA推导的转运蛋白具有多个跨膜结构域,并且在氨基酸序列上与其哺乳动物对应物有一定的相似性。当CPTA和CPTB在非洲爪蟾卵母细胞中表达,并通过放射性示踪剂通量和微电极电压钳技术进行研究时,它们表现出由质子梯度驱动的可饱和的生电转运活性,底物特异性谱较宽且有重叠。这两种转运体都能识别二肽、三肽和四肽,包括苯丙氨酰甲硫氨酰精氨酰苯丙氨酰胺(FMRF酰胺)和N-乙酰天冬氨酰谷氨酸,它们是在整个动物界普遍存在的一个大神经肽家族的成员。然而,动力学分析表明,CPTA和CPTB对肽底物的亲和力不同,前者是高亲和力类型,后者是低亲和力类型。CPTA和CPTB由位于不同染色体上的两个不同基因编码,并在生物体的整个生命周期中表达。

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