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蜂毒肽及其他阳离子试剂对红细胞膜中带3蛋白聚集作用的比较研究

A comparative study of band 3 aggregation in erythrocyte membranes by melittin and other cationic agents.

作者信息

Clague M J, Cherry R J

机构信息

Department of Chemistry and Biological Chemistry, University of Essex, Colchester, U.K.

出版信息

Biochim Biophys Acta. 1989 Mar 27;980(1):93-9. doi: 10.1016/0005-2736(89)90204-6.

Abstract

The technique of laser flash-induced transient dichroism has been used to measure the rotational diffusion of eosin-labelled band 3 proteins in erythrocyte ghosts. A retardation in the mobility of band 3, measured subsequent to the addition of a variety of polyvalent cationic species, has been interpreted to reflect aggregation or 'clustering' of the protein in the plane of the membrane. A comparative study is reported between three such aggregators: melittin, polylysine and Zn2+, wherein their respective abilities to induce aggregation have been measured under varying conditions. Unlike that for melittin, band 3 aggregation by polylysine and Zn2+ is shown to be sensitive to proteolytic degradation of the membrane and to the ionic strength of the surrounding medium. Studies with fragments of melittin derived from its chymotryptic cleavage show the hydrophilic C-terminal 20-26 section to possess independent aggregating ability, but also the requirement of the 1-19 hydrophobic section to be attached in order to prevent reversibility by high ionic strength buffers. Melittin is also shown to have a unique ability to aggregate bacteriorhodopsin reconstituted into DMPC vesicles, which is partially retained by its 1-19 but not by its 20-26 fragment.

摘要

激光闪光诱导瞬态二色性技术已被用于测量红细胞血影中嗜酸性标记的带3蛋白的旋转扩散。在添加各种多价阳离子后测量带3迁移率的延迟,被解释为反映了膜平面中蛋白质的聚集或“簇集”。本文报道了三种这样的聚集剂:蜂毒肽、聚赖氨酸和Zn2+之间的比较研究,其中它们各自诱导聚集的能力已在不同条件下进行了测量。与蜂毒肽不同,聚赖氨酸和Zn2+诱导的带3聚集对膜的蛋白水解降解和周围介质的离子强度敏感。对蜂毒肽经胰凝乳蛋白酶切割产生的片段进行的研究表明,亲水性的C末端20 - 26片段具有独立的聚集能力,但也需要连接1 - 19疏水片段以防止高离子强度缓冲液使其逆转。蜂毒肽还显示出具有独特的能力,能够聚集重组到二肉豆蔻酰磷脂酰胆碱(DMPC)囊泡中的细菌视紫红质,其1 - 19片段部分保留了这种能力,而20 - 26片段则没有。

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