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Oligosaccharide specificity of influenza H1N1 virus neuraminidases

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Summary

A fluorescent neuraminidase (NA) assay has been developed; 20 samples in five replicates could be analyzed at the same time, allowing us to study the kinetics of the enzyme-substrate interaction. The specificities of six influenza H1N1 virus NAs for BODIPY-labeled 3′SiaLac, 3′SiaLacNAc, SiaLec, SiaLea, 6′SiaLac, and 6′SiaLacNAc were evaluated. The duck virus NA hydrolyzed 6′SiaLac and 6′SiaLacNAc 50 times more slowly than 2–3 isomers. Swine viruses digested SiaLea and 2–6 sialosides 20 times more slowly than 2–3 trisaccharides. For the human viruses, the difference between 2–6 and 2–3 oligosaccharides desialylation efficiency did not exceed five times; notably, the inner core of 2–3 sialosaccharide was discriminated. The results are evidence that influenza virus NAs can distinguish substrate structure at the tri- and tetrasaccharide level.

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Abbreviations

BODIPY (FLSE):

4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid (succinimidyl ester)

DRM:

diluted reaction mixture

HAU:

hemagglutinating unit

K M :

Michaelis constant

Lac:

Galβ1-4Glc

NA:

neuraminidase

Neu5Ac:

N-acetylneuraminic acid

S 0 :

initial substrate concentration

TBA:

thiobarbituric acid

V 0 and V max :

initial and maximal rate of enzymatic hydrolysis, respectively

References

  • JF Barros SuffixJr DS Alviano MH Silva MD Wigg CS Alviano R Shauer JNSS Couceiro (2003) ArticleTitleCharacterization of sialidase from an influenza A (H3N2) virus strain: kinetic parameters and substrate specificity Intervirology 46 199–206 Occurrence Handle10.1159/000072428 Occurrence Handle1:CAS:528:DC%2BD3sXmsFaqtLc%3D

    Article  CAS  Google Scholar 

  • LG Baum JC Paulson (1991) ArticleTitleThe N2 neuraminidase of human influenza virus has acquired a substrate specificity complementary to the hemagglutinin receptor specificity Virology 180 10–15 Occurrence Handle1984642 Occurrence Handle10.1016/0042-6822(91)90003-T Occurrence Handle1:CAS:528:DyaK3MXnslGrug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • IV Berezin AA Klesov (1975) Praktichesky kurs himicheskoy i fermentativnoy kinetiki Mosow University Moscow

    Google Scholar 

  • Berezin IV, Martinek K (1977) Basics of physical chemistry of enzymatic catalysis, High School, Moscow

  • RA Copeland (2000) Enzymes: a practical introduction to structure, mechanism, and data analysis Wiley-VCH New York

    Google Scholar 

  • JNSS Couceiro LJ Baum (1994) ArticleTitleCharacterization of the hemagglutinin receptor specificity and neuraminidase substrate specificity of clinical isolates of human influenza A viruses Mem Inst Oswaldo Cruz Rio de Janeiro 89 587–591 Occurrence Handle1:STN:280:BymD2czotFA%3D

    CAS  Google Scholar 

  • A Fersht (1985) Enzyme structure and mechanism W.H. Freeman New York

    Google Scholar 

  • MN Janakiraman CL White WG Laver GM Air M Luo (1994) ArticleTitleStructure of influenza virus neuraminidase B/Lee/40 complexed with sialic acid and dehydro analog at 1.8-Å resolution: implications for the catalytic mechanism Biochemistry 33 8172–8179 Occurrence Handle8031750 Occurrence Handle10.1021/bi00193a002 Occurrence Handle1:CAS:528:DyaK2cXksFejt78%3D

    Article  PubMed  CAS  Google Scholar 

  • D Katinger L Mochalova A Chinarev N Bovin J Romanova (2004) ArticleTitleSpecificity of neuraminidase activity from influenza viruses isolated in different hosts tested with novel substrates Arch Virol 149 2131–2140 Occurrence Handle15503202 Occurrence Handle10.1007/s00705-004-0364-1 Occurrence Handle1:CAS:528:DC%2BD2cXoslGisbk%3D

    Article  PubMed  CAS  Google Scholar 

  • D Kobasa S Kodihalli M Luo MR Castrucci I Donatelli Y Suzuki T Suzuki Y Kawaoka (1999) ArticleTitleAmino acid resides contributing to the substrate specificity of the influenza A virus neuraminidase J Virol 73 6743–6751 Occurrence Handle10400772 Occurrence Handle1:CAS:528:DyaK1MXksFKnuro%3D

    PubMed  CAS  Google Scholar 

  • D Kobasa K Wells Y Kawaoka (2001) ArticleTitleAmino acids responsible for the absolute sialidase activity of the influenza virus neuraminidase: relationship to growth in the duck intestine J Virol 75 11773–117780 Occurrence Handle11689658 Occurrence Handle10.1128/JVI.75.23.11773-11780.2001 Occurrence Handle1:CAS:528:DC%2BD3MXotlaqsLo%3D

    Article  PubMed  CAS  Google Scholar 

  • J McKimm-Breschkin (2000) ArticleTitleResistance of influenza viruses to neuraminidase inhibitors – a review Antivir Res 47 1–17 Occurrence Handle10930642 Occurrence Handle10.1016/S0166-3542(00)00103-0 Occurrence Handle1:CAS:528:DC%2BD3cXltlWgsrk%3D

    Article  PubMed  CAS  Google Scholar 

  • LV Mochalova EY Korchagina VS Kurova JA Styria AS Gambaryan NV Bovin (2005) ArticleTitleFluorescent assay for studying the substrate specificity of neuraminidase Anal Biochem 34 190–193 Occurrence Handle10.1016/j.ab.2005.02.019 Occurrence Handle1:CAS:528:DC%2BD2MXjslKjtr0%3D

    Article  CAS  Google Scholar 

  • G Pazynina A Tuzikov A Chinarev P Obukhova N Bovin (2002) ArticleTitleSimple stereoselective synthesis of alfa2-6 sialooligosaccharides Tetrahedron Lett 43 8011–8013 Occurrence Handle10.1016/S0040-4039(02)01983-4 Occurrence Handle1:CAS:528:DC%2BD38XnvVymt74%3D

    Article  CAS  Google Scholar 

  • GV Pazynina MA Sablina AB Tuzikov AA Chinarev NV Bovin (2003) ArticleTitleSynthesis of complex 2–3 sialooligosaccharides, including sulfated and fucosylated ones, using Neu5Ac alfa2-3Gal as a building block Mendeleev Commun 13 245–248 Occurrence Handle10.1070/MC2003v013n06ABEH001845 Occurrence Handle1:CAS:528:DC%2BD2cXpsVWjug%3D%3D

    Article  CAS  Google Scholar 

  • J Romanova D Katinger B Ferko R Voglauer L Mochalova N Bovin W Lim H Katinger A Egorov (2003) ArticleTitleDistinct host range of influenza H3N2 virus isolates in Vero and MDCK cells is determined by cell specific glycosylation pattern Virology 307 90–97 Occurrence Handle12667817 Occurrence Handle10.1016/S0042-6822(02)00064-8 Occurrence Handle1:CAS:528:DC%2BD3sXitlWnt74%3D

    Article  PubMed  CAS  Google Scholar 

  • S Sabesan S Neira F Davidson J Duus K Bock (1994) ArticleTitleSynthesis and enzymatic and NMR studies of novel sialoside probes: unprecedented, selective neuraminidase hydrolysis of and inhibition by C-6-(methyl)-Gal sialosides J Am Chem Soc 116 1616–1634 Occurrence Handle10.1021/ja00084a003 Occurrence Handle1:CAS:528:DyaK2cXhvFaitL4%3D

    Article  CAS  Google Scholar 

  • AB Tuzikov AS Gambaryan LR Juneja NV Bovin (2000) ArticleTitleConversion of complex sialooligosaccharides into polymeric conjugates and their anti-influenza virus inhibitory potency J Carbohyd Chem 19 1191–1200 Occurrence Handle1:CAS:528:DC%2BD3MXivF2ksg%3D%3D Occurrence Handle10.1080/07328300008544143

    Article  CAS  Google Scholar 

  • L Warren (1959) ArticleTitleThe thiobarbituric acid assay of sialic acids J Biol Chem 234 1971–1975 Occurrence Handle13672998 Occurrence Handle1:CAS:528:DyaG1MXhtFamtrk%3D

    PubMed  CAS  Google Scholar 

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Mochalova, L., Kurova, V., Shtyrya, Y. et al. Oligosaccharide specificity of influenza H1N1 virus neuraminidases. Arch Virol 152, 2047–2057 (2007). https://doi.org/10.1007/s00705-007-1024-z

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  • DOI: https://doi.org/10.1007/s00705-007-1024-z

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