Functional homologs of cyanovirin-N amenable to mass production in prokaryotic and eukaryotic hosts.
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Mori T, Barrientos LG, Han Z, Gronenborn AM, Turpin JA, Boyd MR
Functional homologs of cyanovirin-N amenable to mass production in prokaryotic and eukaryotic hosts.
Protein Expr Purif. 2002 Oct;26(1):42-9.
- PubMed ID
- 12356469 [ View in PubMed]
- Abstract
Cyanovirin-N (CV-N) is under development as a topical (vaginal or rectal) microbicide to prevent sexual transmission of human immunodeficiency virus (HIV), and an economically feasible means for very large-scale production of the protein is an urgent priority. We observed that N-glycosylation of CV-N in yeast eliminated the anti-HIV activity, and that dimeric forms and aggregates of CV-N occurred under certain conditions, potentially complicating the efficient, large-scale manufacture of pure monomeric CV-N. We therefore expressed and tested CV-N homologs in which the glycosylation-susceptible Asn residue at position 30 was replaced with Ala, Gln, or Val, and/or the Pro at position 51 was replaced by Gly to eliminate potential conformational heterogeneity. All homologs exhibited anti-HIV activity comparable to wild-type CV-N, and the Pro51Gly homologs were significantly more stable proteins. These glycosylation-resistant, functional cyanovirins should be amenable to large-scale production either in bacteria or in eukaryotic hosts.