Inhibition of trypsin by cowpea thionin: characterization, molecular modeling, and docking.
Higher plants produce several families of proteins with toxic properties, which act as defense compounds against pests and pathogens. The thionin family represents one family and comprises low molecular mass cysteine-rich proteins, usually basic and distributed in different plant tissues. Here, we report the purification and characterization of a new thionin from cowpea (Vigna unguiculata) with proteinase inhibitory activity. Cowpea thionin inhibits trypsin, but not chymotrypsin, binding with a stoichiometry of 1:1 as shown with the use of mass spectrometry. Previous annotations of thionins as proteinase inhibitors were based on their erroneous identification as homologues of Bowman-Birk family inhibitors. Molecular modeling experiments were used to propose a mode of docking of cowpea thionin with trypsin. Consideration of the dynamic properties of the cowpea thionin was essential to arrive at a model with favorable interface characteristics comparable with structures of trypsin-inhibitor complexes determined by X-ray crystallography. In the final model, Lys11 occupies the S1 specificity pocket of trypsin as part of a canonical style interaction.
Melo
Francislete R
FR
Departamento de Biologia Celular, Universidade de Brasília, Brasília-DF, Brasil. etemelo@cenargen.embrapa.br
Rigden
Daniel J
DJ
Franco
Octávio L
OL
Mello
Luciane V
LV
Ary
Maria B
MB
Grossi de Sá
Maria F
MF
Bloch
Carlos
C
D000595:Amino Acid Sequence; D001665:Binding Sites; D003198:Computer Simulation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding; D016415:Sequence Alignment; D014357:Trypsin / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D010940:Plant Proteins; D014361:Trypsin Inhibitors; D014357:Trypsin