Bacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide

Arpan Deb, William A. Johnson, Alexander P. Kline, Boston J. Scott, Lydia R. Meador, Dustin Srinivas, Jose M. Martin-Garcia, Katerina Dörner, Chad Borges, Rajeev Misra, Brenda Hogue, Petra Fromme, Tsafrir Leket-Mor

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Viral 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.

Original languageEnglish (US)
Article numbere0172529
JournalPLoS One
Volume12
Issue number2
DOIs
StatePublished - Feb 1 2017

Fingerprint

viral proteins
Viral Proteins
Protein Sorting Signals
Human immunodeficiency virus 1
signal peptide
membrane proteins
pectate lyase
HIV-1
Membrane Proteins
Proteins
Membranes
Pectobacterium carotovorum
circular dichroism spectroscopy
CD4 Antigens
Structural Models
light scattering
Dynamic light scattering
Cell membranes
Circular Dichroism
Viruses

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Bacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide. / Deb, Arpan; Johnson, William A.; Kline, Alexander P.; Scott, Boston J.; Meador, Lydia R.; Srinivas, Dustin; Martin-Garcia, Jose M.; Dörner, Katerina; Borges, Chad; Misra, Rajeev; Hogue, Brenda; Fromme, Petra; Leket-Mor, Tsafrir.

In: PLoS One, Vol. 12, No. 2, e0172529, 01.02.2017.

Research output: Contribution to journalArticle

Deb, Arpan ; Johnson, William A. ; Kline, Alexander P. ; Scott, Boston J. ; Meador, Lydia R. ; Srinivas, Dustin ; Martin-Garcia, Jose M. ; Dörner, Katerina ; Borges, Chad ; Misra, Rajeev ; Hogue, Brenda ; Fromme, Petra ; Leket-Mor, Tsafrir. / Bacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide. In: PLoS One. 2017 ; Vol. 12, No. 2.
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AU - Meador, Lydia R.

AU - Srinivas, Dustin

AU - Martin-Garcia, Jose M.

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