Regulation of receptor binding affinity of influenza virus hemagglutinin by its carbohydrate moiety

The hemagglutinin (HA) of the fowl plague virus (FPV) strain of influenza A virus has two N-linked oligosaccharides attached to Asn123 and Asn149 in the vicinity of the receptor binding site. The effect of these carbohydrate side chains on the binding of HA to neuraminic acid-containing receptors has been analyzed. When the oligosaccharides were deleted by site-specific mutagenesis, HA expressed from a simian virus 40 vector showed enhanced hemadsorbing activity. Binding was so strong under these conditions that erythrocytes were no longer released by viral neuraminidase and that release was significantly reduced when neuraminidase from Vibrio cholerae was used. Similarly, when these oligosaccharides were removed selectively from purified viruses by N-glycosidase F, such virions were unable to elute from receptors, although they retained neuraminidase activity. Thus, release of FPV from cell receptors depends on the presence of the HA glycans at Asn123 and Asn149. On the other hand, receptor binding was abolished when these oligosaccharides were sialylated after expression in the absence of neuraminidase (M. Ohuchi, A. Feldmann, R. Ohuchi, and H.-D. Klenk, Virology 212:77-83, 1995). These observations indicate that the receptor affinity of FPV HA is controlled by oligosaccharides adjacent to the receptor binding site.

[1]  J. Skehel,et al.  Structure of influenza haemagglutinin at the pH of membrane fusion , 1994, Nature.

[2]  I. Schulze,et al.  Single amino acid substitutions in the hemagglutinin can alter the host range and receptor binding properties of H1 strains of influenza A virus , 1991, Journal of virology.

[3]  H. Klenk,et al.  Mutations at the cleavage site of the hemagglutinin after the pathogenicity of influenza virus A/chick/Penn/83 (H5N2). , 1989, Virology.

[4]  J. Mermod,et al.  Increased biological activity of deglycosylated recombinant human granulocyte/macrophage colony-stimulating factor produced by yeast or animal cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[5]  H. Klenk,et al.  Host cell proteases controlling virus pathogenicity. , 1994, Trends in microbiology.

[6]  A. Varki,et al.  Biological roles of oligosaccharides: all of the theories are correct , 1993, Glycobiology.

[7]  M. Namiki,et al.  Purification and characterization of three forms of differently glycosylated recombinant human granulocyte-macrophage colony-stimulating factor. , 1991, Archives of biochemistry and biophysics.

[8]  J. Oxford,et al.  Structural changes in the haemagglutinin which accompany egg adaptation of an influenza A(H1N1) virus. , 1987, Virology.

[9]  I. Wilson,et al.  Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution , 1981, Nature.

[10]  H. Klenk,et al.  Carbohydrate masking of an antigenic epitope of influenza virus haemagglutinin independent of oligosaccharide size. , 1992, Glycobiology.

[11]  C. Naeve,et al.  Alterations in the hemagglutinin associated with adaptation of influenza B virus to growth in eggs. , 1985, Virology.

[12]  R. Webster,et al.  Glycosylation affects cleavage of an H5N2 influenza virus hemagglutinin and regulates virulence. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Webster,et al.  Direct sequencing of the HA gene of influenza (H3N2) virus in original clinical samples reveals sequence identity with mammalian cell-grown virus , 1990, Journal of virology.

[14]  A. Burgess,et al.  Granulocyte-macrophage colony stimulating factor from human lymphocytes. The effect of glycosylation on receptor binding and biological activity. , 1990, The Journal of biological chemistry.

[15]  R. Dwek,et al.  Effects of N-glycosylation on in vitro activity of Bowes melanoma and human colon fibroblast derived tissue plasminogen activator. , 1989, Biochemistry.

[16]  G. Larsen,et al.  Functional effects of asparagine-linked oligosaccharide on natural and variant human tissue-type plasminogen activator. , 1988, The Journal of biological chemistry.

[17]  H. Klenk,et al.  Neuraminidase is essential for fowl plague virus hemagglutinin to show hemagglutinating activity. , 1995, Virology.

[18]  H. Klenk,et al.  Human influenza virus hemagglutinin with high sensitivity to proteolytic activation , 1991, Journal of virology.

[19]  H. Klenk,et al.  Rescue of vector-expressed fowl plague virus hemagglutinin in biologically active form by acidotropic agents and coexpressed M2 protein , 1994, Journal of virology.

[20]  A. Wittwer,et al.  Glycosylation at Asn-184 inhibits the conversion of single-chain to two-chain tissue-type plasminogen activator by plasmin. , 1990, Biochemistry.

[21]  H. Klenk,et al.  Oligosaccharides in the stem region maintain the influenza virus hemagglutinin in the metastable form required for fusion activity , 1997, Journal of virology.

[22]  A. Wittwer,et al.  Oligosaccharides at each glycosylation site make structure-dependent contributions to biological properties of human tissue plasminogen activator. , 1991, Glycobiology.

[23]  R. Webster,et al.  Extensive heterogeneity in the hemagglutinin of egg-grown influenza viruses from different patients. , 1989, Virology.

[24]  R. Lamb,et al.  Influenza virus M2 protein ion channel activity stabilizes the native form of fowl plague virus hemagglutinin during intracellular transport , 1994, Journal of virology.

[25]  H. Klenk,et al.  Carbohydrates of influenza virus. Structural elucidation of the individual glycans of the FPV hemagglutinin by two‐dimensional 1H n.m.r. and methylation analysis. , 1985, The EMBO journal.

[26]  R. Dwek,et al.  N-glycosylation and in vitro enzymatic activity of human recombinant tissue plasminogen activator expressed in Chinese hamster ovary cells and a murine cell line. , 1989, Biochemistry.

[27]  J. Oxford,et al.  A host cell selected variant of influenza B virus with a single nucleotide substitution in HA affecting a potential glycosylation site was attenuated in virulence for volunteers , 1988 .

[28]  J. Sambrook,et al.  Cell-free synthesis of enzymically active tissue-type plasminogen activator. Protein folding determines the extent of N-linked glycosylation. , 1992, The Biochemical journal.

[29]  J. Wood,et al.  Sequence analysis of the haemagglutinin (HA) of influenza A (H1N1) viruses present in clinical material and comparison with the HA of laboratory-derived virus. , 1991, The Journal of general virology.

[30]  I. Wilson,et al.  A carbohydrate side chain on hemagglutinins of Hong Kong influenza viruses inhibits recognition by a monoclonal antibody. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[31]  P. Hung,et al.  Deglycosylation Increases the Fibrinolytic Activity of a Deletion Mutant of Tissue-Type Plasminogen Activator , 1990, Thrombosis and Haemostasis.

[32]  W. Garten,et al.  A H1 hemagglutinin of a human influenza A virus with a carbohydrate-modulated receptor binding site and an unusual cleavage site , 1993, Virus Research.