Oligosaccharide specificity of influenza H1N1 virus neuraminidases.
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, SiaLe(c), SiaLe(a), 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 SiaLe(a) 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.
Russian Academy of Sciences, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
L
L
Mochalova
V
V
Kurova
Y
Y
Shtyrya
E
E
Korchagina
A
A
Gambaryan
I
I
Belyanchikov
N
N
Bovin
D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D004372:Ducks / Q000821:virology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000201:enzymology*; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000821:virology; D007700:Kinetics; D009439:Neuraminidase / Q000378:metabolism*; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism*; D009976:Orthomyxoviridae Infections / Q000821:virology; D013379:Substrate Specificity; D013552:Swine / Q000821:virology
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D009844:Oligosaccharides; D009439:Neuraminidase