Summary
The acetylesterase of influenza C virus has been reported recently to be inhibited by diisopropylfluorophosphate (DFP) [Muchmore EA, Varki A (1987) Science 236: 1293–1295]. As this inhibitor is known to bind covalently to the serine in the active site of serine esterases, we attempted to determine the serine in the active site of the influenza C acetylesterase. Incubation of purified influenza C virus with3H-DFP resulted in the selective labelling of the influenza C glycoprotein HEF. The labelled glycoprotein was isolated from a SDS-polyacrylamide gel. Following reduction and carboxymethylation, tryptic peptides of HEF were prepared and analyzed by reversed phase HPLC. The peptide containing the3H-DFP was subjected to sequence analysis. The amino acids determined from the NH2-terminus were used to locate the peptide on the HEF polypeptide. Radiosequencing revealed that3H-DFP is attached to amino acid 17 of the tryptic peptide. These results indicate that serine 71 is the active-site serine of the acetylesterase of influenza C virus.
Article PDF
Avoid common mistakes on your manuscript.
References
Bradshaw RA, Cancedda F, Ericsson LH, Neumann PA, Piccoli SP, Schlesinger MJ, Shriefer K, Walsh KA (1981) Amino acid sequence of Escherichia coli alkaline phosphatase. Proc Natl Acad Sci USA 78: 3473–3477
Buonagurio DA, Nakada S, Desselberger U, Krystal M, Palese P (1985) Noncumulative sequence changes in the hemagglutinin genes of influenza C virus isolates. Virology 146: 221–232
Formanowski F, Meier-Ewert H (1988) Isolation of the influenza C glycoprotein in a soluble form by bromelain digestion. Virus Res 10: 177–192
Herrler G, Compans RW, Meier-Ewert H (1979) A precursor glycoprotein in influenza C virus. Virology 99: 49–56
Herrler G, Nagele A, Meier-Ewert H, Bhown AS, Compans RW (1981) Isolation and structural analysis of influenza C virion glycoproteins. Virology 113: 439–451
Herrler G, Geyer R, Müller H-P, Stirm S, Klenk H-D (1985) Rat α1-macroglobulin inhibits hemagglutination by influenza C virus. Virus Res 2: 183–192
Herrler G, Rott R, Klenk H-D, Müller H-P. Shukla AK, Schauer R (1985) The receptor-destroying enzyme of influenza C virus is neuraminate O-acetylesterase. EMBO J 4: 1503–1506
Herrler G, Klenk H-D (1987) The surface receptor is a major determinant of the cell tropism of influenza C virus. Virology 159: 102–108
Herrler G, Dürkop I, Becht H, Klenk H-D (1988) The glycoprotein of influenza C virus is the haemagglutinin, esterase and fusion factor. J Gen Virol 69: 839–846
Kitame F, Sugawara K, Ohwada K, Homma M (1982) Proteolytic activation of hemolysis and fusion by influenza C virus. Arch Virol 73: 357–361
MacPhee-Quigley K, Taylor P, Taylor S (1985) Primary structure of the catalytic subunits from two molecular forms of acetylcholinesterase. J Biol Chem 260: 12185–12189
Meier-Ewert H, Nagele A, Herrler G, Basak S, Compans RW (1981) Structure of the spike glycoprotein of influenza C virus. In: Nayak D, Fox CD (eds) Genetic variation among influenza A viruses. Academic Press, New York, pp 263–272
Muchmore EA, Varki A (1987) Selective inactivation of influenza C esterase: a probe for detecting 9-O-acetylated sialic acids. Science 236: 1293–1295
Nakada S, Creager RS, Krystal M, Aaronson RP, Palese P (1984) Influenza C virus hemagglutinin: comparison with influenza A and B virus hemagglutinins. J Virol 50: 118–124
Ohuchi M, Ohuchi R, Mifune K (1982) Demonstration of hemolytic and fusion activities of influenza C virus. J Virol 42: 1076–1079
Pfeifer JB, Compans RW (1984) Structure of the infuenza C glycoprotein as determined from cloned DNA. Virus Res 1: 281–296
Rogers GN, Herrler G, Paulson JC, Klenk H-D (1986) Influenza C virus uses 9-O-acetyl-N-acetylneuraminic acid as a high affinity receptor determinant for attachment to cells. J Biol Chem 261: 5947–5951
Vlasak R, Krystal M, Nacht M, Palese P (1987) The influenza C virus glycoprotein (HE) exhibits receptor-binding (hemagglutinin) and receptor-destroying activities. Virology 160: 419–425
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Herrler, G., Multhaup, G., Beyreuther, K. et al. Serine 71 of the glycoprotein HEF is located at the active site of the acetylesterase of influenza C virus. Archives of Virology 102, 269–274 (1988). https://doi.org/10.1007/BF01310831
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01310831