Gene structure of the 'large' sialidase isoenzyme from Clostridium perfringens A99 and its relationship with other clostridial nanH proteins.
Clostridium perfringens possesses two sialidase isoenzymes of different molecular weight. Almost 90% of the gene encoding the 'large' form was found on a 3.1 kb chromosomal fragment (Sau3AI) of strain A99 by hybridization with probes developed from the N-terminal protein sequence and from commonly conserved sialidase motifs ('Asp-boxes'), whereas the remaining 3'-terminal part was detected on a 2.1 kb fragment (Hind III) of chromosomal DNA. After combination of both fragments, the resulting E. coli clones expressed sialidase activity, the properties of the recombinant sialidase corresponding with those of the wild type enzyme. The entire chromosomal fragment of 3665 bp encompasses the complete sialidase gene of 2082 bp corresponding to 694 amino acids, from which a molecular weight of 72,956 for the mature protein can be deduced. The first 41 amino acids are mostly hydrophobic and probably represent a signal peptide. The sialidase structural gene follows a non-coding region with an inverted repeat and a ribosome-binding site. Upstream from the regulatory region, another open reading frame (ORF) was detected. The 3'-terminus of the sialidase structural gene is directly followed by a further ORF of unknown function, which possibly encodes a putative permease or the acylneuraminate pyruvate-lyase involved in sialic acid catabolism. The primary structure of the 'large' isoenzyme is very similar to the sialidase of Clostridium septicum (55% identical amino acids), whereas the homology with the 'small' form of the same species is comparatively low (26%).
Traving
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Biochemisches Institut der Christian-Albrechts-Universität, Kiel.
Schauer
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Roggentin
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D000595:Amino Acid Sequence; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology / Q000235:genetics; D003016:Clostridium perfringens / Q000201:enzymology* / Q000235:genetics*; D004269:DNA, Bacterial / Q000378:metabolism; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000235:genetics* / Q000378:metabolism; D015345:Oligonucleotide Probes; D010802:Phylogeny; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D007527:Isoenzymes; D015345:Oligonucleotide Probes; D009439:Neuraminidase