Purification And Characterization Of A New Trypsin Inhibitor From Dimorphandra Mollis Seeds

A second trypsin inhibitor (DMTI-II) was purified from the seed of Dimorphandra mollis (Leguminosae-Mimosoideae) by ammonium sulfate precipitation (30-60%), gel filtration, and ionexchange and affinity chromatography. A molecular weight of 23 kDa was estimated by gel filtration on a Superdex 75 column SDS-PAGE under reduced conditions showed that DMTI-II consisted of a single polypeptide chain, although isoelectric focusing revealed the presence of three isoforms. The dissociation constant of 1.7 ́ 10-9 M with bovine trypsin indicated a high affinity between the inhibitor and this enzyme. The inhibitory activity was stable over a wide pH range and in the presence of DTT. The N-terminal sequence of DMTI-II showed a high degree of homology with other Kunitz-type inhibitors. ©2002 Plenum Publishing Corporation.208625632Batista, I.F.C., Oliva, M.L.V., Araujo, M.S., Sampaio, M.U., Richardson, M., Fritz, H., Sampaio, C.A.M., (1996) Phytochemistry, 41, pp. 1017-1022Bernfeld, P., (1955) Methods in Enzymology, 1, 149p. , Collowick, S. P. and Kaplan, N. O., eds. Academic Press, New YorkBirk, Y., (1994) Arch. Latinoam. Nutricion, 44, pp. 26S-30SBradford, M.M., (1970) Anal. Biochem., 72, pp. 248-254Broze, G.J., Girard, T.J., Novotny, W.F., (1990) Biochemistry, 29, pp. 7539-7546DeClerck, Y.A., Imren, S., (1994) Eur. J. Cancer, 30 A, pp. 2170-2180Di Ciero, L., Oliva, M.L.V., Torquato, R., Köhler, P., Weder, J.K.P., Novelle, J.C., Sampaio, C.A.M., Marangoni, S., (1998) J. Protein. Chem., 17, pp. 827-834Erlanger, B.F., Kolowsky, N., Cohen, N., (1961) Arch. Biochem. Biophys., 95, pp. 271-278Heath, R.L., McDonald, G., Christeller, J.T., Lee, M., Bateman, K., West, J., Heeswijck, R.V., Anderson, M.A., (1997) J. Insect Physiol., 43, pp. 833-842Jongsma, M., Bolter, C., (1997) J. Insect Physiol., 43, pp. 885-895Joulbert, F.J., Carlsson, F.H., Haylett, T., (1981) Hoppe-Seyler's Zeitsch. Physiol. Chem., 362, pp. 531-538Kalume, D.F., Sousa, M.V., Morhy, L., (1995) J. Protein Chem., 14, pp. 685-693Kennedy, A.R., (1998) Pharmacol. Ther., 78, pp. 167-209Knight, C.G., (1986) Proteinase Inhibitors, pp. 23-51. , Barret, A. J. and Salvesen, G., eds., Eisevier, AmsterdamKunitz, M., (1945) J. Gen. Physiol., 30, pp. 291-310Laemmli, U.K., (1970) Nature, 227, pp. 680-685Lehle, K., Kohnert, U., Stern, A., Popp, F., Jaenicke, R., (1996) Nature Biotech., 14, pp. 476-480Lourenço, D.M., Sampaio, M.U., Sampaio, C.A.M., (1989) Adv. Med. Exp. Biol., 247 B, pp. 553-557Macedo, M.L.R., Matos, D.G.G., Machado, O.L.T., Marangoni, S., Novello, J.C., (2000) Phytochemistry, 54, pp. 553-558Macedo, M.L.R., Xavier-Filho, J., (1992) J. Sci. Food. Agric., 58, pp. 55-58Mosolov, V.V., (1995) Appl. Biochem. Microbiol., 31, pp. 3-7Norioka, N., Hara, S., Ikenaka, T., Abe, J., (1988) Agric. Biol. Chem., 52, pp. 1245-1252Odani, S., Ono, T., Ikenaka, T., (1979) J. Biochem., 86, pp. 1795-1805Oliva, M.L.V., Grisolla, D., Sampaio, M.U., Sampaio, C.A.M., (1982) Agents Actions, 9, pp. 52-57Oliva, M.L.V., Mendes, C.R., Juliano, M.A., Chagas, J.R., Rosa, J.C., Greene, L.J., Sampaio, M.U., Sampaio, C.A.M., (1999) Immunopharmacology, 45, pp. 163-169Oliva, M.L.V., Sallai, R.C., Sampaio, C.A.M., Fritz, H., Auerswald, E.A., Tanaka, A.S., Sampaio, M.U., (1996) Immunopharmacology, 32, pp. 85-87Oliva, M.L.V., Santomauro-Vaz, E.M., Andrade, S.A., Juliano, M.A., Pott, V.J., Sampaio, M.U., Sampaio, C.A.M., (2001) Biol. Chem., 382, pp. 109-113Oliva, M.L.V., Souza-Pinto, J.C., Batista, I.F.C., Araujo, M.S., Silveira, V.F., Auerswald, E.A., Mentele, R., Sampaio, C.A.M., (2000) Biochim. Biophys. Acta, 1477, pp. 64-74Ramasarma, P.R., Rao, A.G.A., Rao, D.R., (1995) Biochim. Biophys. Acta, 1248, pp. 35-42Richardson, M., (1991) Methods in Plant Biochemistry, 5. , Dey, P. M. and Harborne, J. B., eds., Academic Press, New YorkRyan, C.A., (1990) Anna. Rev. Phytopathol., 28, pp. 425-449Sampaio, C.A.M., Oliva, M.L.V., Sampaio, M.U., Batista, I.F.C., Bueno, N.R., Tanaka, A.S., Auerswald, A., Fritz, H., (1996) Itntnunopharmacology, 32, pp. 62-66Schuler, T.H., Poppy, G.M., Kerry, B.R., Denholm, I., (1998) Trends Biotechnol., 16, pp. 168-175Shewry, P.R., Lucas, J.A., (1997) Adv. Botan. Res., 26, pp. 135-192Souza, E.M.T., Mizuta, K., Sampaio, M.U., Sampaio, C.A.M., (1995) Phytochemistry, 39, pp. 521-525Uriel, J., Berges, J., (1968) Nature, 218, pp. 578-580Valueva, T.A., Mosolov, V.V., (1999) J. Plant Physiol., 46, pp. 322-329Valueva, T.A., Revina, T.A., Mosolov, V.V., (1997) Biochemistry, 62, pp. 1367-1374. , Translated from BiokhimiyaWalsh, T.A., Twitchell, W.P., (1991) Plant Phys., 97, pp. 15-18Westermeir, R., (1993) In Electrophoresis in Practice-A Guide to Theory and Practice, pp. 210-212. , VCH Publishers, New YorkXavier-Filho, J., (1992) Rev. Bras. Fisiol. Veg., 4, pp. 1-6Xavier-Filho, J., Campos, F.A.P., (1983) Brazilian J. Med. Biol. Res., 16, pp. 11-15Xavier-Filho, J., Campos, F.A.P., Ary, M.B., Silva, C.P., Carvalho, M.M.M., Macedo, M.L.R., Lemos, F.J.A., Grant, G., (1989) J. Agric. Food Chem., 37, pp. 1139-114

Análise de trilha em caracteres produtivos de Pennisetum sob corte em Itambé, Pernambuco Path analysis in productive characters of Pennisetum under cut in Itambé-PE

Objetivou-se obter estimativas de coeficientes de correlação genética e avaliar os desdobramentos das correlações em efeitos diretos e indiretos (análise de trilha) das variáveis independentes explicativas sobre a produção de matéria seca (variável dependente principal) em cinco clones de capim-elefante. Foram realizados cinco cortes a cada 60 dias nas condições edafoclimáticas da Zona da Mata de Pernambuco. Cada parcela experimental apresentou 24 m², com 2 m² de área útil. Houve alta correlação entre quase todas as variáveis explicativas e a variável principal. Entretanto, a característica número de lâminas foliares por perfilho explicou melhor o potencial de produção de matéria seca e atuou de forma direta e indireta sobre as variáveis explicativas.<br>This work aimed to estimate the correlation coefficients, determining the direct and indirect effects (path analysis) of explanatory independent variables on total dry matter production (main dependent variables) of five elephant grass clones in five cuts accomplished at the forest zone of Pernambuco. Each experimental plot measured 24 m² with 2 m² of utile area. There was high correlation between almost all the independent variables and the main variable. The characteristic leaf blade number per tiller, however, explained better the potential of dry matter production and leaf blade dry matter production, acting, respectively, in a direct and indirect way on the explanatory variables