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    Characterisation of multiple trypsins from the midgut of Locusta migratoria

    Lam, W. and Coast, Geoffrey and Rayne, Richard (2000) Characterisation of multiple trypsins from the midgut of Locusta migratoria. Insect Biochemistry and Molecular Biology 30 (1), pp. 85-94. ISSN 0965-1748.

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    Abstract

    Three isoforms of trypsin were identified in midgut preparations from Locusta migratoria. Ammonium-sulphate-fractionated luminal contents of midguts were subjected to benzamidine affinity chromatography; proteins eluted by benzamidine were then separated by anion-exchange chromatography. Cationic (TRY 1) and anionic (TRY 2) trypsin activities were eluted from the DEAE column. TRY 1 was homogeneous, producing a single band of Mr 23,000 on SDS-PAGE. TRY 2 comprised two trypsins, TRY 2A (Mr 27,000) and TRY 2B (Mr 29,000). Following a subsequent chromatography step using a Bio-Rad UNO Q column, TRY 2A and TRY 2B were resolved to homogeneity. When homogenates of midgut caecae were the starting material for chromatography, SDS-PAGE of benzamidine-eluted proteins revealed an additional putative trypsin of Mr 17,000 (termed SERP 17) which had been absent from luminal enzyme preparations. Determination of the N-terminal 11 amino acid residues of each protein revealed unique, but similar sequences. The four sequences all began with IVGG, a motif which signifies all four proteins are serine proteases. TRY 1, TRY 2A and TRY 2B were shown to contain only trypsin activity and the preparations were sensitive to inhibition by AEBSF, PMSF, TLCK, benzamidine, leupeptin, SBTI, BPTI and E64.

    Metadata

    Item Type: Article
    School: Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences
    Depositing User: Sarah Hall
    Date Deposited: 14 May 2019 09:31
    Last Modified: 02 Aug 2023 17:51
    URI: https://eprints.bbk.ac.uk/id/eprint/27515

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