Analysis of a peptide inhibitor of paramyxovirus (NDV) fusion using biological assays, NMR, and molecular modeling.
To investigate the molecular mechanisms involved in paramyxovirus-induced cell fusion, the function and structure of a peptide with a 20-amino-acid sequence from the leucine zipper region (heptad repeat region 2) of the Newcastle disease virus fusion protein (F) were characterized. A peptide with the sequence ALDKLEESNSKLDKVNVKLT (amino acids 478-497 of the F protein) was found to inhibit syncytia formation after virus infection and after transfection of Cos cells with the HN (hemagglutinin-neuraminidase) and F protein cDNAs. Using an F protein gene that requires addition of exogenous trypsin for cleavage, it was shown that the peptide exerted its inhibitory effect prior to cleavage. The three-dimensional conformation of the peptide in aqueous solution was determined through the use of NMR and molecular modeling. Results showed that the peptide formed a helix with properties between an alpha-helix and a 3(10)-helix and that leucine residues aligned along one face of the helix. Side chain salt bridges and hydrogen bonds likely contributed to the stability of the peptide secondary structure. Analysis of the aqueous solution conformation of the peptide suggested mechanisms for specificity of interaction with the intact F protein.
Young
J K
JK
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical Center, Worcester 01655, USA.
Hicks
R P
RP
Wright
G E
GE
Morrison
T G
TG
D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D015707:HN Protein / Q000235:genetics / Q000378:metabolism; D016350:Leucine Zippers; D008561:Membrane Fusion / Q000187:drug effects*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009522:Newcastle disease virus / Q000187:drug effects* / Q000502:physiology; D019906:Nuclear Magnetic Resonance, Biomolecular; D010452:Peptide Biosynthesis; D010446:Peptide Fragments; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D017433:Protein Structure, Secondary; D013696:Temperature; D013997:Time Factors; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000502:physiology
D000577:Amides; D015707:HN Protein; D010446:Peptide Fragments; D010455:Peptides; D014760:Viral Fusion Proteins