Effects of a novel pathogenesis-related class 10 (PR-10) protein from Crotalaria pallida Roots with papain inhibitory activity against root-knot nematode Meloidogyne incognita.
A novel pathogenesis-related class 10 (PR-10) protein with papain inhibitory activity, named CpPRI, was purified from Crotalaria pallida roots by ammonium sulfate precipitation followed by three reverse-phase high-performance liquid chromatographies (HPLCs). CpPRI is made up of a single polypeptide chain with a M(r) of 15 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This protein exhibited a K(i) value of 1.8 x 10(-9) M and operates via a noncompetitive inhibition mechanism. The alignment of the N-terminal amino acid sequence of CpPRI with other proteins revealed its identity with PR-10 proteins. CpPRI acts against digestive proteinase from root-knot nematode Meloidogyne incognita and demonstrated nematostatic and nematicide effects on this parasite in bioassays. In a localization study, fluorescein-5-isothiocyanate (FITC)-CpPRI was observed to internalize and diffuse over the entire J2 body after 6 h of incubation. This fact could explain the natural tolerance of this plant species to nematodes.
Andrade
Lúcia Betânia da S
LB
Centro de Ciencias Agrarias e Biologicas, Universidade Estadual Vale do Acarau (UVA), Sobral, Ceara (CE), Brazil.
Oliveira
Adeliana S
AS
Ribeiro
Jannison K C
JK
Kiyota
Sumika
S
Vasconcelos
Ilka Maria
IM
de Oliveira
José Tadeu Abreu
JT
de Sales
Maurício P
MP
D000818:Animals; D029864:Crotalaria / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D007700:Kinetics; D018551:Solanum lycopersicum / Q000469:parasitology; D008970:Molecular Weight; D010206:Papain / Q000037:antagonists & inhibitors*; D010935:Plant Diseases / Q000469:parasitology*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D018517:Plant Roots / Q000469:parasitology; D014431:Tylenchoidea / Q000187:drug effects* / Q000502:physiology
D004791:Enzyme Inhibitors; D010940:Plant Proteins; D010206:Papain