Construction, expression, and analysis of recombinant HIV gp41 constructs containing a novel cellular binding domain

Daniel Kestler, L. A. Henderson, J. D. Noti

Research output: Contribution to journalArticle

Abstract

The gp41 polypeptide of human immunodeficiency virus (HIV) contains an immunosuppressive domain, an epitope which elicits specific cytolytic T cell responses to HIV, and a complement Clq interactive domain. In addition, a synthetic peptide called CS3, derived from gp41 (amino acids 576–593 of gp160) and contiguous with the major immunodominant domain, binds to cellular proteins and may be important in HIV entry/fusion. In order to further investigate the role of the CS3 region of gp41 in cellular binding and to investigate other properties of gp41, sufficient quantities of this polypeptide must be readily available. We have therefore cloned the region of the HIV genome between nucleotides 7891 and 8188 (corresponding to amino acids 541–639 of gp160) into a series of procaryotic expression vectors. The resulting clones express a recombinant polypeptide of gp41 (r41). Two of these recombinants, pMAL‐cRl/r41 and pGEMEX‐2/r41, expressed the highest and most consistent levels of r41 as judged by both sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and Western blot analysis. With the pMAL‐cRl/r41 construct, r41 was expressed as a fusion to the maltose‐binding protein (MBP) and, following purification by affinity chromatography, was cleaved from MBP by factor Xa protease digestion. MBP/r41 may be useful for studies of a reported gp41 cellular binding domain and may facilitate studies involving other functions ascribed to this region of gp41. © 1993 John Wiley & Sons, Inc.

Original languageEnglish (US)
Pages (from-to)81-86
Number of pages6
JournalBiotechnology and Bioengineering
Volume42
Issue number1
DOIs
StatePublished - Jan 1 1993
Externally publishedYes

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Viruses
Polypeptides
HIV
Proteins
Peptides
Amino acids
Fusion reactions
Epitopes
Affinity chromatography
Amino Acids
Immunodominant Epitopes
Virus Internalization
Factor Xa
T-cells
Immunosuppressive Agents
Nucleotides
Electrophoresis
Affinity Chromatography
Purification
Digestion

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Construction, expression, and analysis of recombinant HIV gp41 constructs containing a novel cellular binding domain. / Kestler, Daniel; Henderson, L. A.; Noti, J. D.

In: Biotechnology and Bioengineering, Vol. 42, No. 1, 01.01.1993, p. 81-86.

Research output: Contribution to journalArticle

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