MEMBRANE BIOGENESIS AND MEMBRANE FUSION

膜生物发生和膜融合

• 批准号: 3291043
• 负责人: ANTHONY W SCOTTO
• 金额: $ 8.99万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1990-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3291043
• 关键词: electron microscopy; laboratory rat; liposomes; membrane fusion; membrane model; membrane proteins; membrane reconstitution /synthesis; membrane structure; phospholipids; protein biosynthesis; vesicle /vacuole

The object of this proposal is to continue the study of two aspects of our recent discovery in the field of membrane biology. The first, is the spontaneous insertion of integral membrane proteins into preformed bilayers and the second is the membrane fusion that is catalyzed by the same methodology. We have observed that the presence of impurities in the bilayer of preformed vesicles triggers both the incorporation of purified integral membrane proteins as well as the fusion of unilamellar proteoliposomes when the phospholipids are in the gel state. The rapid fusion of liposomes and proteoliposomes by this manipulation of the phospholipid bilayer presents an ideal model system for the study of the cellular membrane fusion at the level of the phospholipid bilayer. We propose to utilize this system to investigate several concepts in membrane biology while we extend the applicability of this method for reconstruction studies. We plan to construct proteoliposomes with several different membrane proteins by this procedure; some of the integral membrane proteins for this study will be selected to demonstrate a functional interaction within the newly formed membrane while others will be employed to modify the surface availability and characteristics of the proteoliposome. As a result of this aspect of the work, we expect to be able to construct a model cell membrane composed of several different integral membrane proteins in which many of the current concepts in cellular and membrane biology can be tested. Furthermore, the apparent ability to construct proteoliposomes of cellular dimension (7 microns) by this procedure will be further examined in an effort to design and construct a model cell of predetermined protein composition and lipid/protein ratio. The stability of these very large proteoliposomes of high lipid/protein ratio has exceeded our expectations and we believe that the proposed proteoliposomes of low lipid-to-protein ratio will provide a very stable vesicle which may represent the ideal model cell. The long range goals are two-fold with respect to the phenomenon we have discovered. We propose to determine whether the mechanism by which we have incorporated integral membrane proteins into preformed bilayers represents the in vivo participation of the phospholipid in cellular membranes. We will determine too whether the specificity of membrane fusion among biological membrane is regulated in part by the proposed mechanism for the fusion of proteoliposomes in this procedure.

这项建议的目的是继续研究我们的两个方面， 膜生物学领域的最新发现。 第一个是 完整膜蛋白自发插入预先形成的双层 第二种是由相同的催化剂催化的膜融合 方法论 我们已经观察到， 预先形成的囊泡的双层触发了纯化的 整合膜蛋白以及单层细胞的融合 当磷脂处于凝胶状态时，蛋白脂质体。 快速 脂质体和脂蛋白体的融合 磷脂双分子层是研究细胞膜结构的理想模型系统。 磷脂双分子层水平的细胞膜融合。 我们 我建议利用这个系统来研究膜中的几个概念 生物学，同时我们扩展了这种方法的适用性， 问题研究 我们计划用几种不同的蛋白脂质体 膜蛋白;一些完整的膜蛋白 将选择用于本研究以证明功能性相互作用 在新形成的膜内，而其他的将被用来改变 蛋白脂质体的表面可用性和特性。 作为 由于这方面的工作，我们希望能够建立一个 由几种不同的完整膜组成的模型细胞膜 蛋白质，其中许多目前的概念，在细胞和膜 生物学是可以检验的。 此外，明显的能力， 将通过该方法制备细胞尺寸（7微米）的蛋白脂质体。 进一步研究，努力设计和构建一个模型细胞， 预定的蛋白质组成和脂质/蛋白质比率。 稳定性 这些非常大的高脂质/蛋白质比的蛋白脂质体中， 超出了我们的预期，我们相信， 低的脂蛋白比将提供非常稳定的囊泡， 代表理想的模型细胞。 长期目标是双重的， 我们发现的现象。 我们建议确定 我们整合膜的机制 蛋白质进入预先形成的双层代表了在体内参与 细胞膜中的磷脂。 我们也将确定 生物膜之间膜融合的特异性受到调节， 部分由蛋白脂质体融合的拟议机制， procedure.

项目成果

Spontaneous incorporation of bacteriorhodopsin into large preformed vesicles.

• DOI: 10.1021/bi00483a012
• 发表时间: 1990-08
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: A. Scotto;M. Gompper