SYNTHESIS AND DISTRIBUTION OF PROTEINS IN MEMBRANES

蛋白质在膜中的合成和分布

• 批准号: 6725397
• 负责人: DAVID D SABATINI
• 金额: $ 48.94万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6725397
• 关键词: Golgi apparatus; MDCK cell; acyltransferase; cell free system; enzyme activity; fatty acid binding protein; immunoelectron microscopy; intracellular membranes; intracellular transport; laboratory mouse; laboratory rabbit; laboratory rat; lysophospholipids; membrane activity; membrane biogenesis; membrane proteins; phosphatidylinositols; phospholipase D; protein biosynthesis; protein localization; protein purification; protein transport; transport proteins; vesicle /vacuole

DESCRIPTION (Verbatim from the applicant's abstract): This application is concerned with the nature of the carrier vesicles that mediate the transport of proteins from the trans-Golgi network (TGN) to the cell surface of epithelial cells. It proposes to study the process of their formation and to establish the mode of action of the specific proteins, lipids and cofactors that in the cytosol and TGN membranes play essential roles in this process. For these studies we use a cell free system that we developed that recreates in vitro the generation of post-Golgi vesicles from a purified Golgi fraction obtained from virus-infected MDCK cells that contains accumulated sialylated VSV-G protein molecules. For vesicle generation, the Golgi fraction is incubated with cytosolic proteins and a source of nucleoside triphosphates. The vesicles, when produced in the presence of GTPgammaS, contain a coatomer coat. They also contain rab11, which has been previously localized to the TGN and implicated in transport of VSV-G to the plasma membrane through the endosomal compartment The formation of post-Golgi vesicles can be effected in two sequential phases, a first one of Arf-dependent coat assembly/bud formation and a second one of membrane scission. We will identify the cytosolic proteins and lipid that play a key role in the scission of post-Golgi vesicles and determine their role in the process. These include the phospatidylinositol transfer protein (PITP), a scission factor of unknown protein composition that contains bourn lysophospatidic acid (LPA) and is likely to be a lysophosphatidic acid acyltransferase, (possibly BARS-50, the target of the Golgi tubulating agent, BFA), a fatty acid binding protein (FABP) that keeps the scission factor inactive and a postulated factor that restricts cleavage of coatomer-coated membranes to the necks of coated buds. We will test a model in which activated Arf sets the scission machinery in motion at the neck of a coated bud by dissociating the acyltransferase from the FABP that keeps it inactive. We will elucidate the possible role in scission of a phospholipase D and that of a PKC-like molecule that regulates it. We will also identify protein molecules in the coat and membranes of the TGN-derived that contribute to their targetting to an acceptor compartment and to identify the nature of that compartment. Since the vesicles contain rab11, we will determine whether the vesicle can dock and fuse with endosomes.

描述（逐字摘自申请人摘要）：本申请是

项目成果

Nucleotide sequence of rat preputial gland beta-glucuronidase cDNA and in vitro insertion of its encoded polypeptide into microsomal membranes.

大鼠包皮腺 β-葡萄糖醛酸酶 cDNA 的核苷酸序列及其编码的多肽在体外插入微粒体膜。

• DOI: 10.1073/pnas.83.19.7292
• 发表时间: 1986
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Nishimura,Y;Rosenfeld,MG;Kreibich,G;Gubler,U;Sabatini,DD;Adesnik,M;Andy,R
• 通讯作者: Andy,R

Biosynthesis and intracellular sorting of growth hormone-viral envelope glycoprotein hybrids.

生长激素-病毒包膜糖蛋白杂合体的生物合成和细胞内分选。

• DOI: 10.1083/jcb.101.4.1351
• 发表时间: 1985
• 期刊: The Journal of cell biology
• 作者: Rizzolo,LJ;Finidori,J;Gonzalez,A;Arpin,M;Ivanov,IE;Adesnik,M;Sabatini,DD
• 通讯作者: Sabatini,DD

Regulation of post-Golgi vesicle production in an in vitro system.

体外系统中高尔基体后囊泡产生的调节。

• DOI: 10.1101/sqb.1995.060.01.021
• 发表时间: 1995
• 期刊: Cold Spring Harbor symposia on quantitative biology
• 作者: Simon,JP;Ivanov,IE;Ren,M;Zeng,J;Shopsin,B;Hersh,D;Tempst,P;Erdjument-Bromage,H;Lui,M;DeLemos-Chiarandini,C
• 通讯作者: DeLemos-Chiarandini,C

Functional protein prenylation is required for the brefeldin A-dependent retrograde transport from the Golgi apparatus to the endoplasmic reticulum.

功能性蛋白质异戊二烯化是布雷菲德菌素 A 依赖性从高尔基体逆行转运至内质网所必需的。

• DOI: 10.1074/jbc.272.33.20828
• 发表时间: 1997
• 期刊: The Journal of biological chemistry
• 作者: Ivessa,NE;Gravotta,D;DeLemos-Chiarandini,C;Kreibich,G
• 通讯作者: Kreibich,G

Coatomer, but not P200/myosin II, is required for the in vitro formation of trans-Golgi network-derived vesicles containing the envelope glycoprotein of vesicular stomatitis virus.

• DOI: 10.1073/pnas.95.3.1073
• 发表时间: 1998-02
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: J. Simon;T. H. Shen;I. Ivanov;D. Gravotta;T. Morimoto;M. Adesnik;D. Sabatini