BIROn - Birkbeck Institutional Research Online

    Characterisation of multiple trypsins from the midgut of locusta migratoria

    Lam, W. and Coast, G.M. 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.

    Full text not available from this repository.

    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: School of Science > Biological Sciences
    Depositing User: Sarah Hall
    Date Deposited: 29 Apr 2019 16:23
    Last Modified: 29 Apr 2019 16:23
    URI: https://eprints.bbk.ac.uk/id/eprint/27355

    Statistics

    Downloads
    Activity Overview
    0Downloads
    220Hits

    Additional statistics are available via IRStats2.

    Archive Staff Only (login required)

    Edit/View Item Edit/View Item