Analysis by phage display selection and site-directed retromutagenesis of the Mustard Trypsin Inhibitor 2 reactive site

Abstract

The Mustard Trypsin Inhibitor (MSI) family is a small family of plant protease inhibitors so far onlyfound in Brassicaceae. Using a phage display selection, MTI-2 (Mustard Trypsin Inhibitor 2) mutants weredetected and analysed for their biochemical characteristics. Retromutants of the selected MTI-2 proteinswere constructed and expressed in the Pichia pastoris system. The recombinant proteins were analysedby activity assays against bovine trypsin and Helicoverpa zea trypsin, and by circular dichroism. Theseanalyses suggest a strict requirement for a specific proline residue adjacent to the inhibitor reactive siteand give additional insights for future phage display application.


Autore Pugliese

Tutti gli autori

  • Volpicella M.; Leoni C.; Arnesano F.; Gallerani R.; Ceci L.R.

Titolo volume/Rivista

Journal of plant physiology


Anno di pubblicazione

2010

ISSN

0176-1617

ISBN

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Nessuna citazione

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