Surface-exposed Amino Acid Residues of HPV16 L1 Protein Mediating Interaction with Cell Surface Heparan Sulfate
| Autor: | Hans-Christoph Selinka, Xiaojiang S. Chen, Rolf E. Streeck, Martin Sapp, Dorothe Spillmann, Maren Knappe, Ulf Lindahl, Sabrina Bodevin |
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| Rok vydání: | 2007 |
| Předmět: |
Surface Properties
Lysine Plasma protein binding Biochemistry Antibodies chemistry.chemical_compound Capsid Chlorocebus aethiops Animals Humans Computer Simulation Molecular Biology chemistry.chemical_classification Alanine Infectivity Heparin Cell Membrane Capsomere Oncogene Proteins Viral Cell Biology Heparan sulfate Amino acid chemistry Mutagenesis COS Cells Capsid Proteins Heparitin Sulfate Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 282:27913-27922 |
| ISSN: | 0021-9258 |
| Popis: | Efficient infection of cells by human papillomaviruses (HPVs) and pseudovirions requires primary interaction with cell surface proteoglycans with apparent preference for species carrying heparan sulfate (HS) side chains. To identify residues contributing to virus/cell interaction, we performed point mutational analysis of the HPV16 major capsid protein, L1, targeting surface-exposed amino acid residues. Replacement of lysine residues 278, 356, or 361 for alanine reduced cell binding and infectivity of pseudovirions. Various combinations of these amino acid exchanges further decreased cell attachment and infectivity with residual infectivity of less than 5% for the triple mutant, suggesting that these lysine residues cooperate in HS binding. Single, double, or triple exchanges for arginine did not impair infectivity, demonstrating that interaction is dependent on charge distribution rather than sequence-specific. The lysine residues are located within a pocket on the capsomere surface, which was previously proposed as the putative receptor binding site. Fab fragments of binding-neutralizing antibody H16.56E that recognize an epitope directly adjacent to lysine residues strongly reduced HS-mediated cell binding, further corroborating our findings. In contrast, mutation of basic surface residues located in the cleft between capsomeres outside this pocket did not significantly reduce interaction with HS or resulted in assembly-deficient proteins. Computer-simulated heparin docking suggested that all three lysine residues can form hydrogen bonds with 2-O-, 6-O-, and N-sulfate groups of a single HS molecule with a minimal saccharide domain length of eight monomer units. This prediction was experimentally confirmed in binding experiments using capsid protein, heparin molecules of defined length, and sulfate group modifications. |
| Databáze: | OpenAIRE |
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