Fachbereich Veterinärmedizin



    Influenza virus hemagglutinin spike neck architectures and interaction with model enzymes evaluated by MALDI-TOF mass spectrometry and bioinformatics tools (2011)

    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Serebryakova, Marina V
    Kordyukova, Larisa V
    Semashko, Tatiana A
    Ksenofontov, Alexander L
    Rudneva, Irina A
    Kropotkina, Ekaterina A
    Filippova, Irina Yu
    Veit, Michael (WE 5)
    Baratova, Lyudmila A
    Virus research; 160(1-2) — S. 294–304
    ISSN: 0168-1702
    DOI: 10.1016/j.virusres.2011.07.002
    Pubmed: 21763731
    Institut für Virologie

    Robert-von-Ostertag-Str. 7-13
    Gebäude 35
    14163 Berlin
    +49 30 838 51833

    Abstract / Zusammenfassung

    Interactions between model enzymes and the influenza virus hemagglutinin (HA) homotrimeric spike were addressed. We digested influenza virions (naturally occurring strains and laboratory reassortants) with bromelain or subtilisin Carlsberg and analyzed by MALDI-TOF mass spectrometry the resulting HA2 C-terminal segments. All cleavage sites, together with (minor) sites detected in undigested HAs, were situated in the linker region that connects the transmembrane domain to the ectodomain. In addition to cleavage at highly favorable amino acids, various alternative enzyme preferences were found that strongly depended on the HA subtype/type. We also evaluated the surface electrostatic potentials, binding cleft topographies and spatial dimensions of stem bromelain (homologically modeled) and subtilisin Carlsberg (X-ray resolved). The results show that the enzymes (∼45Å(3)) would hardly fit into the small (∼18-20Å) linker region of the HA-spike. However, the HA membrane proximal ectodomain region was predicted to be intrinsically disordered. We propose that its motions allow steric adjustment of the enzymes' active sites to the neck of the HA spike. The subtype/type-specific architectures in this region also influenced significantly the cleavage preferences of the enzymes.