Enzymatic glycosylation of triazole-linked GlcNAc/Glc-peptides: synthesis, stability and anti-HIV activity of triazole-linked HIV-1 gp41 glycopeptide C34 analogues.
Long-lasting sweet proteins: The chemoenzymatic synthesis of a triazole (T)-linked glycosylated C34 fragment from HIV-1 gp41 is described. The glycopeptide shows high solubility, excellent fusion inhibition, and as shown in the graph, promising protease resistance. Endoglycosidase-catalyzed transglycosylation of triazole-linked glucose (Glc) and N-acetylglucosamine (GlcNAc)-containing dipeptides and polypeptides was achieved by using synthetic sugar oxazoline as the donor substrate. It was found that both N- and C-linked Glc/GlcNAc-containing triazole derivatives were effective substrates for endo-beta-N-acetylglucosaminidase from Arthrobacter (Endo-A) for transglycosylation; this demonstrates a broad acceptor substrate specificity for Endo-A. This chemoenzymatic method was successfully used for the synthesis of a novel triazole-linked C34 glycopeptide derived from the HIV-1 envelope glycoprotein, gp41. We found that the synthetic C34 glycopeptide possesses potent anti-HIV activity with an IC(50) of 21 nM. The triazole-linked C34 glycopeptide demonstrated a much enhanced stability against protease- and glycoamidase-catalyzed digestion; this shows the protective effects of glycosylation and the stability of the triazole linkage. These favorable properties suggest that the triazole-linked C34 glycopeptide might be valuable for further development as an anti-HIV drug candidate.
Huang
Wei
W
Institute of Human Virology, University of Maryland, School of Medicine, Baltimore, MD 21201, USA.
Groothuys
Stan
S
Heredia
Alonso
A
Kuijpers
Brian H M
BH
Rutjes
Floris P J T
FP
van Delft
Floris L
FL
Wang
Lai-Xi
LX
D000595:Amino Acid Sequence; D019380:Anti-HIV Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D001173:Arthrobacter / Q000201:enzymology; D055162:Biocatalysis; D002240:Carbohydrate Sequence; D004355:Drug Stability; D006020:Glycopeptides / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D006031:Glycosylation; D015700:HIV Envelope Protein gp41 / Q000737:chemistry*; D020128:Inhibitory Concentration 50; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D014230:Triazoles / Q000737:chemistry*
D019380:Anti-HIV Agents; D006020:Glycopeptides; D015700:HIV Envelope Protein gp41; D014230:Triazoles; C514787:gp41 protein, Human immunodeficiency virus 1; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase