Protein Design of a Bacterially Expressed HIV-1 gp41 Fusion Inhibitor†

Yiqun Deng,‡ Qi Zheng,‡ Thomas J. Ketas,§ John P. Moore,§ and Min Lu*‡
Department of Biochemistry and Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, New York 10021
Biochemistry, 2007, 46 (14), pp 4360–4369
DOI: 10.1021/bi7001289
Publication Date (Web): March 20, 2007
Copyright © 2007 American Chemical Society
†

 This work was supported by NIH Grants U19 AI65413 and R01 AI68591 and by the Irma T. Hirschl Trust. The Department of Microbiology and Immunology at the Weill Medical College gratefully acknowledges the support of the William Randolph Hearst Foundation.

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‡

 Department of Biochemistry.

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§

 Department of Microbiology and Immunology.

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*

 To whom correspondence should be addressed. Phone:  (212) 746-6562. Fax:  (212) 746-8875. E-mail:  mlu@med.cornell.edu.

Abstract

Abstract Image

Peptides derived from the carboxyl-terminal heptad repeat of the gp41 envelope glycoprotein ectodomain (C-peptides) can inhibit HIV-1 membrane fusion by binding to the amino-terminal trimeric coiled coil of the same protein. The fusion inhibitory peptide T-20 contains an additional tryptophan-rich sequence motif whose binding site extends beyond the gp41 coiled-coil region yet provides the key determinant of inhibitory activity in T-20. Here we report the design of a recombinant peptide inhibitor (called C52L) that includes both the C-peptide and tryptophan-rich regions. By calorimetry, C52L binds to a peptide mimic of the amino-terminal coiled coil with a Kd of 80 nM, reflecting the large degree of helicity in C52L as measured by circular dichroism spectroscopy. The C52L peptide potently inhibits in vitro infection of human T cells by diverse primary HIV-1 isolates irrespective of coreceptor preference, with nanomolar IC50 values. Significantly, C52L is fully active against T-20-resistant variants in a single-cycle HIV-1 infectivity assay. Moreover, because it can be expressed in bacteria, the C52L peptide might be more economical to manufacture on a large scale than T-20-like peptides produced by chemical synthesis. Hence the C52L fusion inhibitor may find a practical application, for example as a vaginal or rectal microbicide to prevent HIV-1 infection in the developing world.

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History

  • Published In Issue April 10, 2007
  • Received January 22, 2007
    Revised Manuscript Received February 13, 2007

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