Linker-extended native cyanovirin-N facilitates PEGylation and potently inhibits HIV-1 by targeting the glycan ligand.

Abstract

Cyanovirin-N (CVN) potently inhibits human immunodeficiency virus type 1 (HIV-1) infection, but both cytotoxicity and immunogenicity have hindered the translation of this protein into a viable therapeutic. A molecular docking analysis suggested that up to 12 residues were involved in the interaction of the reverse parallel CVN dimer with the oligosaccharide targets, among which Leu-1 was the most prominent hot spot residue. This finding provided a possible explanation for the lack of anti-HIV-1 activity observed with N-terminal PEGylated CVN. Therefore, linker-CVN (LCVN) was designed as a CVN derivative with a flexible and hydrophilic linker (Gly4Ser)3 at the N-terminus. The N-terminal α-amine of LCVN was PEGylated to create 10 K PEG-aldehyde (ALD)-LCVN. LCVN and 10 K PEG-ALD-LCVN retained the specificity and affinity of CVN for high mannose N-glycans. Moreover, LCVN exhibited significant anti-HIV-1 activity with attenuated cytotoxicity in the HaCaT keratinocyte cell line and MT-4 T lymphocyte cell lines. 10 K PEG-ALD-LCVN also efficiently inactivated HIV-1 with remarkably decreased cytotoxicity and pronounced cell-to-cell fusion inhibitory activity in vitro. The linker-extended CVN and the mono-PEGylated derivative were determined to be promising candidates for the development of an anti-HIV-1 agent. This derivatization approach provided a model for the PEGylation of biologic candidates without introducing point mutations.

• Authors: Jia Chen, Dane Huang, Wei Chen, Chaowan Guo, Bo Wei, Chongchao Wu, Zhou Peng, Jun Fan, Zhibo Hou, Yongsheng Fang, Yifei Wang, Kaio Kitazato, Guoying Yu, Chunbin Zou, Chuiwen Qian, Sheng Xiong
• Journal: PloS one (PLoS One) Vol. 9 Issue 1 Pg. e86455 ( 2014) ISSN: 1932-6203 [Electronic] United States
• PMID: 24475123 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Proteins
• Carrier Proteins
• Oligosaccharides
• cyanovirin N
• Polyethylene Glycols

Topics

• Bacterial Proteins (chemistry, metabolism, pharmacology)
• Carrier Proteins (chemistry, metabolism, pharmacology)
• Dimerization
• Drug Design
• HIV Infections (drug therapy)
• HIV-1 (drug effects)
• Humans
• Models, Molecular
• Molecular Docking Simulation
• Oligosaccharides (metabolism)
• Polyethylene Glycols (metabolism)
• Protein Engineering (methods)