Evaluation of sequence variability in HIV-1 gp41 C-peptide helix-grafted proteins

Rachel L. Tennyson, Susanne N. Walker, Terumasa Ikeda, Reuben S. Harris, Brian R. McNaughton

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Many therapeutically-relevant protein-protein interactions (PPIs) have been reported that feature a helix and helix-binding cleft at the interface. Given this, different approaches to disrupting such PPIs have been developed. While short peptides (<15 amino acids) typically do not fold into a stable helix, researchers have reported chemical approaches to constraining helix structure. However, these approaches rely on laborious, and often expensive, chemical synthesis and purification. Our premise is that protein-based solutions that stabilize a therapeutically-relevant helix offer a number of advantages. In contrast to chemically constrained helical peptides, or minimal/miniature proteins, which must be synthesized (at great expense and labor), a protein can be expressed in a cellular system (like all current protein therapeutics). If selected properly, the protein scaffold can stabilize the therapeutically-relevant helix. We recently reported a protein engineering strategy, which we call “helix-grafted display” and applied it to the challenge of suppressing HIV entry. We have reported helix-grafted display proteins that inhibit formation of an intramolecular PPI involving HIV gp41 C-peptide helix, and HIV gp41 N-peptide trimer, which contain C-peptide helix-binding clefts. Here, we used yeast display to screen a library of grafted C-peptide helices for N-peptide trimer recognition. Using ‘hits’ from yeast display library screening, we evaluated the effect helix mutations have on structure, expression, stability, function (target recognition), and suppression of HIV entry.

Original languageEnglish (US)
Pages (from-to)1220-1224
Number of pages5
JournalBioorganic and Medicinal Chemistry
Volume26
Issue number6
DOIs
StatePublished - Mar 15 2018

Bibliographical note

Funding Information:
We acknowledge Professor Alan Kennan (Colorado State University) for use of circular dichroism equipment. We also acknowledge Leslie Armstrong ( Colorado State University ) for assistance with flow cytometry. This work was funded, in part, by Colorado State University and the Colorado Center for Drug Discovery (B.R.M). Harris lab studies were supported by NIAID R37 AI064046. RSH is an Investigator of the Howard Hughes Medical Institute.

Funding Information:
We acknowledge Professor Alan Kennan (Colorado State University) for use of circular dichroism equipment. We also acknowledge Leslie Armstrong (Colorado State University) for assistance with flow cytometry. This work was funded, in part, by Colorado State University and the Colorado Center for Drug Discovery (B.R.M). Harris lab studies were supported by NIAID R37 AI064046. RSH is an Investigator of the Howard Hughes Medical Institute.

Publisher Copyright:
© 2017

Keywords

  • HIV
  • Helix
  • Protein engineering
  • Protein evolution
  • gp41

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