The superior folding of a RANTES analogue expressed in lactobacilli as compared to mammalian cells reveals a promising system to screen new RANTES mutants

Massimiliano Secchi, Qiang Xu, Paolo Lusso, Luca Vangelista

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Development of effective topical microbicides for the prevention of HIV-1 sexual transmission represents a primary goal for the control of the AIDS pandemic. The viral coreceptor CCR5, used by the vast majority of primary HIV-1 isolates, is considered a primary target molecule. RANTES and its derivatives are the most suitable protein-based compounds to fight HIV-1 via CCR5 targeting. Yet, receptor activation should be avoided to prevent pro-inflammatory effects and possibly provide anti-inflammatory properties. C1C5 RANTES is a chemokine mutant that exhibits high anti-HIV-1 potency coupled with CCR5 antagonism. However, the need for the formation of an N-terminal intramolecular disulfide bridge between non-natural cysteine residues at positions 1 and 5 represents a challenge for the correct folding of this protein in recombinant expression systems, a crucial step towards its development as a microbicide against HIV-1. We report here a rare case of superior folding in a prokaryote as compared to an eukaryotic expression system. Production of C1C5 RANTES was highly impaired in CHO cells, with a dramatic yield reduction compared to that of wild type RANTES and secretion of the molecule as disulfide-linked dimer. Conversely, a human vaginal isolate of Lactobacillus jensenii engineered to secrete C1C5 RANTES provided efficient delivery of the monomeric protein. This and other reports on successful secretion of complex proteins indicate that lactic acid bacteria are an excellent system for the expression of therapeutic proteins, which can be used as a platform for the engineering of conceptually novel RANTES mutants with potent anti-HIV-1 activity.

Original languageEnglish
Pages (from-to)34-41
Number of pages8
JournalProtein Expression and Purification
Volume68
Issue number1
DOIs
Publication statusPublished - Nov 2009
Externally publishedYes

Fingerprint

Chemokine CCL5
Lactobacillus
HIV-1
Disulfides
Proteins
Local Anti-Infective Agents
CHO Cells
Protein Folding
Pandemics
Anti-Infective Agents
Chemokines
Recombinant Proteins
Cysteine
Lactic Acid
Acquired Immunodeficiency Syndrome
Anti-Inflammatory Agents
Bacteria

Keywords

  • Anti-HIV-1 microbicide
  • Lactobacilli
  • Mammalian cells
  • Protein folding
  • RANTES

ASJC Scopus subject areas

  • Biotechnology

Cite this

The superior folding of a RANTES analogue expressed in lactobacilli as compared to mammalian cells reveals a promising system to screen new RANTES mutants. / Secchi, Massimiliano; Xu, Qiang; Lusso, Paolo; Vangelista, Luca.

In: Protein Expression and Purification, Vol. 68, No. 1, 11.2009, p. 34-41.

Research output: Contribution to journalArticle

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