Bacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide
Johnson, William A.
Kline, Alexander P.
Scott, Boston J.
Meador, Lydia R.
Martin-Garcia, Jose M.
Borges, Chad R.
Hogue, Brenda G.
Mor, Tsafrir S.
AffiliationUniv Arizona, Coll Med Phoenix, Dept Basic Med Sci
MetadataShow full item record
PublisherPUBLIC LIBRARY SCIENCE
CitationBacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide 2017, 12 (2):e0172529 PLOS ONE
Rights© 2017 Deb et al. This is an open access article distributed under the terms of the Creative Commons Attribution License.
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AbstractViral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV-1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/ function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora. The protein was detergent extractable and could be isolated in a very pure form. We demonstrate that the PelB signal peptide successfully targets Vpu to the cell membranes and inserts it as a type I membrane protein. PelB-Vpu was biophysically characterized by circular dichroism and dynamic light scattering experiments and was shown to be an excellent candidate for elucidating structural models.
NoteOpen Access Journal.
VersionFinal published version
SponsorsCenter for Membrane Proteins in Infectious Diseases (MPID) - PSI:Biology program within the National Institute of General Medical Sciences' Protein Structure Initiative (National Institutes of Health grant) [U54GM094599]
Except where otherwise noted, this item's license is described as © 2017 Deb et al. This is an open access article distributed under the terms of the Creative Commons Attribution License.