Cloning, nucleotide sequence, and expression of the Pasteurella haemolytica A1 glycoprotease gene.
Pasteurella haemolytica serotype A1 secretes a glycoprotease which is specific for O-sialoglycoproteins such as glycophorin A. The gene encoding the glycoprotease enzyme has been cloned in the recombinant plasmid pH1, and its nucleotide sequence has been determined. The gene (designated gcp) codes for a protein of 35.2 kDa, and an active enzyme protein of this molecular mass can be observed in Escherichia coli clones carrying pPH1. In vivo labeling of plasmid-encoded proteins in E. coli maxicells demonstrated the expression of a 35-kDa protein from pPH1. The amino-terminal sequence of the heterologously expressed protein corresponds to that predicted from the nucleotide sequence. The glycoprotease is a neutral metalloprotease, and the predicted amino acid sequence of the glycoprotease contains a putative zinc-binding site. The gene shows no significant homology with the genes for other proteases of procaryotic or eucaryotic origin. However, there is substantial homology between gcp and an E. coli gene, orfX, whose product is believed to function in the regulation of macromolecule biosynthesis.
Abdullah
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KM
Guelph-Waterloo Centre for Graduate Work in Chemistry, University of Guelph, Ontario, Canada.
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Mellors
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D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D006023:Glycoproteins / Q000378:metabolism; D007526:Isoelectric Point; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010325:Pasteurella / Q000235:genetics*; D012995:Solubility
D006023:Glycoproteins; D010450:Endopeptidases; D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase