大鼠阴离子交换蛋白合成肽抗体制备及鉴定

Preparation and Analysis of the Antibodies Against Synthetic Peptide Immunologically Related to the Peripheral Domain of Anion Exchanger (Band 3) in Rat Erythrocyte Membrane

根据带 3蛋白 (阴离子交换蛋白 )拓扑学模式、氨基酸保守片段、功能片段、天然抗原表位 ,设计大鼠带 3膜外侧片段合成肽 ,制备抗合成肽 ( 1 2肽 )抗体 ,同时制备抗大片段带 3抗体加以印证 .多项免疫学实验鉴定结果表明 ,该 1 2肽是带 3抗原决定簇之一 ,也是带 3发挥阴离子转运功能的关键肽段 ;1 2肽的氨基酸组成与序列在各种属间高度保守 .免疫金银显色———扫描电镜结果给出该 1 2肽为带 3蛋白膜外侧区片段的最直接证据 .制备的抗带 3抗体可作为研究带 3结构与功能、探讨与带

Based on the conservative sequences, topological, biochemical, physiological and immunological data of anion exchanger (AE, band 3 protein) families, a peptide included 12 amino acids (position 476～487: SKLIKIFQDYPL) was synthesized and the antibodies against this peptide and the native segment of band 3 from rat erythrocyte membrane were prepared. The findings of immunoblotting and anion transport studies indicated that the peptide is an essential segment of the anion transport which is a conservative sequence in mammals (rat, mouse as well as human). Immunoscanning electron microscopic study proved directly that the peptide is located at the peripheral domain of band 3 in rat erythrocyte membrane. The antibodies against band 3 are useful tools in studying the structures and functions of the anion transport protein and investigating the relationship between band 3 and diseases.