title: Specificity of factor Xa in the cleavage of fusion proteins.
abstract: The precursor protein honey bee prepromelittin has been expressed as a fusion protein in Escherichia coli joined to the C-terminus of a truncated form of the bacteriophage gene 10 protein via an engineered recognition sequence for Factor Xa. Factor Xa was found to cleave poorly at the engineered site, giving a low yield of the required prepromelittin. In contrast, cleavage on the C-terminal side of the sequence VLGR at residue 67 in the gene 10 sequence proceeded in high yield. Factor Xa may be inhibited by adjacent hydrophobic sequences on the C-terminal side of a potential cleavage site.
authors.affiliation: Department of Surgery, St. George's Hospital Medical School, Tooting, London, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: He
authors.forename: L
authors.initials: L
authors.lastname: Jin
authors.forename: B
authors.initials: B
authors.lastname: Austen
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000378:metabolism; D001483:Base Sequence; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D015951:Factor Xa / Q000378:metabolism*; D006868:Hydrolysis; D008555:Melitten / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004274:DNA, Recombinant; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D008555:Melitten; C016694:prepromelittin; D015951:Factor Xa