| Texto completo | |
| Autor(es): |
Indarte, Martin
;
Lazza, Cristian M.
;
Assis, Diego
;
Caffini, Nestor O.
;
Juliano, Maria A.
;
Aviles, Francesc X.
;
Daura, Xavier
;
Lopez, Laura M. I.
;
Trejo, Sebastian A.
Número total de Autores: 9
|
| Tipo de documento: | Artigo Científico |
| Fonte: | PLANTA; v. 245, n. 2, p. 11-pg., 2017-02-01. |
| Resumo | |
A new BBI-type protease inhibitor with remarkable structural characteristics was purified, cloned, and sequenced from seeds of Maclura pomifera , a dicotyledonous plant belonging to the Moraceae family. In this work, we report a Bowman-Birk inhibitor (BBI) isolated, purified, cloned, and characterized from Maclura pomifera seeds (MpBBI), the first of this type from a species belonging to Moraceae family. MpBBI was purified to homogeneity by RP-HPLC, total RNA was extracted from seeds of M. pomifera, and the 3'RACE-PCR method was applied to obtain the cDNA, which was cloned and sequenced. Peptide mass fingerprinting (PMF) analysis showed correspondence between the in silico-translated protein and MpBBI, confirming that it corresponds to a new plant protease inhibitor. The obtained cDNA encoded a polypeptide of 65 residues and possesses 10 cysteine residues, with molecular mass of 7379.27, pI 6.10, and extinction molar coefficient of 9105 M-1 cm(-1). MpBBI inhibits strongly trypsin with K (i) in the 10(-10) M range and was stable in a wide array of pH and extreme temperatures. MpBBI comparative modeling was applied to gain insight into its 3D structure and highlighted some distinguishing features: (1) two non-identical loops, (2) loop 1 (CEEESRC) is completely different from any known BBI, and (3) the amount of disulphide bonds is also different from any reported BBI from dicot plants. (AU) | |
| Processo FAPESP: | 12/50191-4 - Sintese,estudo cinetico e aplicacoes de substratos e inibidores de enzimas proteoliticas. |
| Beneficiário: | Maria Aparecida Juliano |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |