Structure-function study of a heptad repeat positioned near the transmembrane domain of Sendai virus fusion protein which blocks virus-cell fusion.
A synthetic heptad repeat, SV-473, derived from Sendai virus fusion protein is a potent inhibitor of virus-cell fusion. In order to understand the mechanism of the inhibitory effect, we synthesized and fluorescently labeled SV-465, an extended version of SV-473 by one more heptad, its mutant peptide A17,24-SV-465, in which two heptadic leucines were substituted with two alanines, and its enatiomer D-SV-465, composed entirely of Damino acids. Similar mutations in the homologous fusion protein of the Newcastle disease virus drastically reduced its activity. The data revealed that SV-465, but not A17,24-SV-465 or its enantiomer, is highly active in inhibiting Sendai virus-induced hemolysis of red blood cells. None of the peptides interfere with the binding of virions to the target red blood cells as demonstrated by hemagglutinin assay. Fluorescence and circular dichroism (CD) spectroscopy indicated that: (i) only SV-465 could self-assemble in aqueous environment; (ii) only SV-465 could co-assemble with two other biologically active heptad repeats derived from Sendai virus fusion protein; (iii) SV-465 has a higher helical content than A17,24-SV-465 in solution, and (iv) all the peptides bind strongly to zwitterionic and negatively charged phospholipids. Polarized attenuated total reflection infrared spectroscopy revealed that they bound as monomers onto the surface of zwitterionic membranes with predominantly alpha-helical structures. The functional role of the amino acid 465-497 domain in Sendai virus-mediated membrane fusion is discussed in light of these findings.
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, 76100 Israel.
J K
JK
Ghosh
S G
SG
Peisajovich
M
M
Ovadia
Y
Y
Shai
D000595:Amino Acid Sequence; D004735:Energy Transfer; D006461:Hemolysis; D006801:Humans; D008081:Liposomes; D008561:Membrane Fusion / Q000187:drug effects*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D010743:Phospholipids; D017433:Protein Structure, Secondary; D020449:Repetitive Sequences, Amino Acid; D018108:Respirovirus; D017550:Spectroscopy, Fourier Transform Infrared; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D014779:Virus Replication
D008081:Liposomes; D008565:Membrane Proteins; D010446:Peptide Fragments; D010743:Phospholipids; D014760:Viral Fusion Proteins