Fatty Acid Acylation of the Fusion Glycoprotein of Human Respiratory Syncytial Virus

• Published in: The Journal of biological chemistry 1989-06, Vol.264 (18), p.10339-10342
• Main Authors: Arumugham, R G, Seid, R C, Doyle, S, Hildreth, S W, Paradiso, P R
• Format: Article
• Language: English
• Subjects: Acylation; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Antigens, Viral - genetics; Antigens, Viral - isolation & purification; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Glycoproteins; HN Protein; Molecular Sequence Data; Palmitic Acid; Palmitic Acids - metabolism; Peptide Fragments - isolation & purification; Proteins; Respiratory Syncytial Viruses - immunology; Respiratory Syncytial Viruses - metabolism; Viral Envelope Proteins; Viral Proteins
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)81623-4

We describe the covalent attachment of palmitate to the fusion glycoprotein of respiratory syncytial virus and the identification of the attachment site. Labeling of respiratory syncytial virus-infected Vero cells with [3H]palmitate, followed by the purification and subsequent analysis of the fusion glycoprotein in conjunction with Polyacrylamide gel electrophoresis, demonstrated that the fatty acid is covalently attached to the F1 subunit of the fusion glycoprotein. The bound palmitate was sensitive to 1 M hydroxylamine at neutral pH. In addition, the release of palmitate label by reduction with sodium borohydride showed that the palmitate is linked to the protein through a thioester bond. Isolation of a radiolabeled peptide from a tryptic digest of the protein and subsequent amino-terminal sequence analysis revealed that the cysteine residue (amino acid residue 550) within the anchor sequence, located at the carboxyl terminus of the F1 subunit, is the covalent attachment site for palmitate.