Intrinsic membrane association of the cytoplasmic tail of influenza virus M2 protein and lateral membrane sorting regulated by cholesterol binding and palmitoylation.
Intrinsic membrane association of the cytoplasmic tail of influenza virus M2 protein and lateral membrane sorting regulated by cholesterol binding and palmitoylation.
复制标题
流感病毒M2蛋白胞质尾部的内在膜关联和胆固醇结合和棕榈酰化调节的侧膜分选
DOI：
10.1042/bj20110706
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Veit M.
中科院分区：
文献类型：
--
作者：
Leventhal;Herrmann;Veit M.
流感病毒跨膜蛋白M2是一个质子通道，但也在新生病毒颗粒从质膜分裂中起作用。M2的CT（细胞质尾部）中的两亲性螺旋应该插入脂质双分子层，从而引起曲率。螺旋的棕榈酰化和通过假定的CRAC（胆固醇识别/相互作用氨基酸共识）基元与胆固醇结合被认为是将M2靶向到筏的边缘，即病毒出芽部位。在本研究中，我们测试了该模型的前提条件，即CT与膜相互作用，酰化和胆固醇结合影响M2的靶向性。M2-CT，纯化为谷胱甘肽转移酶融合蛋白，与[3H]光胆固醇和脂质体相关。CRAC基体中酪氨酸残基的突变阻止了[3H]光胆固醇标记和减少脂质体结合。M2-CT与转染细胞中高尔基体定位的黄色荧光蛋白融合；膜靶向依赖于CRAC和（在较小程度上）棕榈酰化。从表达全长M2-GFP（绿色荧光蛋白）的细胞制备巨质膜囊泡表明，该蛋白部分存在于筏结构域。Raft靶向需要棕榈酰化，但不需要CRAC基序。因此棕榈酰化和胆固醇结合对两亲性螺旋的内在膜结合有不同的影响。
The influenza virus transmembrane protein M2 is a proton channel, but also plays a role in the scission of nascent virus particles from the plasma membrane. An amphiphilic helix in the CT (cytoplasmic tail) of M2 is supposed to insert into the lipid bilayer, thereby inducing curvature. Palmitoylation of the helix and binding to cholesterol via putative CRAC (cholesterol recognition/interaction amino acid consensus) motifs are believed to target M2 to the edge of rafts, the viral-budding site. In the present study, we tested pre-conditions of this model, i.e. that the CT interacts with membranes, and that acylation and cholesterol binding affect targeting of M2. M2-CT, purified as a glutathione transferase fusion protein, associated with [3H]photocholesterol and with liposomes. Mutation of tyrosine residues in the CRAC motifs prevented [3H]photocholesterol labelling and reduced liposome binding. M2-CT fused to the yellow fluorescent protein localized to the Golgi in transfected cells; membrane targeting was dependent on CRAC and (to a lesser extent) on palmitoylation. Preparation of giant plasma membrane vesicles from cells expressing full-length M2–GFP (green fluorescent protein) showed that the protein is partly present in the raft domain. Raft targeting required palmitoylation, but not the CRAC motifs. Thus palmitoylation and cholesterol binding differentially affect the intrinsic membrane binding of the amphiphilic helix.
登录
查看更多内容
影响因子：
2.9
作者：
Nguyen, Phuong A.;Soto, Cinque S.;Polishchuk, Alexei;Caputo, Gregory A.;Tatko, Chad D.;Ma, Chunlong;Ohigashi, Yuki;Pinto, Lawrence H.;DeGrado, William F.;Howard, Kathleen P.
通讯作者：
Howard, Kathleen P.
影响因子：
3.7
作者：
Rossman JS;Lamb RA
通讯作者：
Lamb RA
多碱性区对于流感病毒基质蛋白M1的膜结合不是必需的。
影响因子：
3.7
作者：
B. Thaa;A. Herrmann;M. Veit
通讯作者：
M. Veit
DOI：
10.1073/pnas.98.3.1267
发表时间：
2001-01
影响因子：
11.1
作者：
Hua Li;Z. Yao;Babett Degenhardt;G. Teper;V. Papadopoulos
通讯作者：
Hua Li;Z. Yao;Babett Degenhardt;G. Teper;V. Papadopoulos
DOI：
10.1083/jcb.200412058
发表时间：
2005-06-20
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hess ST;Kumar M;Verma A;Farrington J;Kenworthy A;Zimmerberg J
通讯作者：
Zimmerberg J