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标题：Correlation of the structural and functional domains in the membrane protein Vpu from HIV-1
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作者：F. M. Marassi ; C. Ma ; H. Gratkowski 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1999
卷号：96
期号：25
页码：14336-14341
DOI：10.1073/pnas.96.25.14336
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Vpu is an 81-residue membrane protein encoded by the HIV-1 genome. NMR experiments show that the protein folds into two distinct domains, a transmembrane hydrophobic helix and a cytoplasmic domain with two in-plane amphipathic -helices separated by a linker region. Resonances in one-dimensional solid-state NMR spectra of uniformly 15N labeled Vpu are clearly segregated into two bands at chemical shift frequencies associated with NH bonds in a transmembrane -helix, perpendicular to the membrane surface, and with NH bonds in the cytoplasmic helices parallel to the membrane surface. Solid-state NMR spectra of truncated Vpu2-51 (residues 2-51), which contains the transmembrane -helix and the first amphipathic helix of the cytoplasmic domain, and of a construct Vpu28-81 (residues 28-81), which contains only the cytoplasmic domain, support this structural model of Vpu in the membrane. Full-length Vpu (residues 2-81) forms discrete ion-conducting channels of heterogeneous conductance in lipid bilayers. The most frequent conductances were 22 {+/-} 3 pS and 12 {+/-} 3 pS in 0.5 M KCl and 29 {+/-} 3 pS and 12 {+/-} 3 pS in 0.5 M NaCl. In agreement with the structural model, truncated Vpu2-51, which has the transmembrane helix, forms discrete channels in lipid bilayers, whereas the cytoplasmic domain Vpu28-81, which lacks the transmembrane helix, does not. This finding shows that the channel activity is associated with the transmembrane helical domain. The pattern of channel activity is characteristic of the self-assembly of conductive oligomers in the membrane and is compatible with the structural and functional findings.