Identification and characterization of protease inhibitors in Diplotaxis species.
PCR analysis of the genomes of two wild Brassicaceae plants, Diplotaxis muralis and Diplotaxis tenuifolia, demonstrated the presence of several genes coding for potential protease inhibitors, classifiable within the mustard inhibitor family (MSI). This is a small family of plant protease inhibitors named after the mustard trypsin inhibitor MTI-2, the first protease inhibitor characterized in Brassicaceae. From identified sequences two recombinant inhibitors were expressed in Pichia pastoris. In comparison with MTI-2, they show a reduced activity against bovine trypsin. However, when tested against trypsin-like proteases present in the guts of Helicoverpa zea larvae, the Diplotaxis inhibitors and MTI-2 show similar activities, indicating that the usually adopted procedure of reporting activity of plant protease inhibitors against bovine trypsin may lead to wrong estimation of their effect on insect proteases. This issue is of particular relevance when planning the use of PI genes for developing insect resistant plants.
Volpicella
Mariateresa
M
Department of Biochemistry and Molecular Biology, University of Bari, Via Amendola 165/A, 70126 Bari, Italy.
De Leo
Francesca
F
Sciancalepore
Marta
M
Sonnante
Gabriella
G
Pignone
Domenico
D
Gallerani
Raffaele
R
Ceci
Luigi R
LR
D000595:Amino Acid Sequence; D019607:Brassicaceae / Q000737:chemistry*; D017931:DNA Primers; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011480:Protease Inhibitors / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D011480:Protease Inhibitors