Structural and functional studies on the extracellular domain of BST2/tetherin in reduced and oxidized conformations

HIV-1 and other enveloped viruses can be restricted by a host cellular protein called BST2/tetherin that prevents release of budded viruses from the cell surface. Mature BST2 contains a small cytosolic region, a predicted transmembrane helix, and an extracellular domain with a C-terminal GPI anchor....

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 2010-10, Vol.107 (42), p.17951-17956
Hauptverfasser: Schubert, Heidi L., Zhai, Qianting, Sandrin, Virginie, Eckert, Debra M., Garcia-Maya, Mitla, Saul, Louise, Sundquist, Wesley I., Steiner, Roberto A., Hill, Christopher P., Harrison, Stephen C.
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container_end_page 17956
container_issue 42
container_start_page 17951
container_title Proceedings of the National Academy of Sciences - PNAS
container_volume 107
creator Schubert, Heidi L.
Zhai, Qianting
Sandrin, Virginie
Eckert, Debra M.
Garcia-Maya, Mitla
Saul, Louise
Sundquist, Wesley I.
Steiner, Roberto A.
Hill, Christopher P.
Harrison, Stephen C.
description HIV-1 and other enveloped viruses can be restricted by a host cellular protein called BST2/tetherin that prevents release of budded viruses from the cell surface. Mature BST2 contains a small cytosolic region, a predicted transmembrane helix, and an extracellular domain with a C-terminal GPI anchor. To advance understanding of BST2 function, we have determined a 2.6 Å crystal structure of the extracellular domain of the bacterially expressed recombinant human protein, residues 47–152, under reducing conditions. The structure forms a single long helix that associates as a parallel dimeric coiled coil over its C-terminal two-thirds, while the N-terminal third forms an antiparallel four-helix bundle with another dimer, creating a global tetramer. We also report the 3.45 Å resolution structure of BST2(51-151) prepared by expression as a secreted protein in HEK293T cells. This oxidized construct forms a dimer in the crystal that is superimposable with the reduced protein over the C-terminal two-thirds of the molecule, and its N terminus suggests pronounced flexibility. Hydrodynamic data demonstrated that BST2 formed a stable tetramer under reducing conditions and a dimer when oxidized to form disulfide bonds. A mutation that selectively disrupted the tetramer (L70D) increased protein expression modestly but only reduced antiviral activity by approximately threefold. Our data raise the possibility that BST2 may function as a tetramer at some stage, such as during trafficking, and strongly support a model in which the primary functional state of BST2 is a parallel disulfide-bound coiled coil that displays flexibility toward its N terminus.
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subjects Antigens, CD - chemistry
Antigens, CD - metabolism
Biochemistry
Biological Sciences
Biopolymers - chemistry
Cells
Chemical bonds
Crystal structure
Crystallography, X-Ray
Crystals
Dimers
Disulfides
Epithelial cells
GPI-Linked Proteins - chemistry
GPI-Linked Proteins - metabolism
HIV 1
Humans
Membranes
Molecules
Mutation
Oxidation
Oxidation-Reduction
Protein Conformation
Proteins
Structure-Activity Relationship
Virions
Viruses
title Structural and functional studies on the extracellular domain of BST2/tetherin in reduced and oxidized conformations
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