Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel α‐amylase inhibitory properties

Physiologia Plantarum - Tập 142 Số 3 - Trang 233-246 - 2011
Mariângela S. S. Diz1, André de Oliveira Carvalho1, Suzanna F. F. Ribeiro1, Maura Da Cunha2, Leila Maria Beltramini3, Rosana Rodrigues4, Viviane Veiga do Nascimento1, Olga Lima Tavares Machado5, Valdirene Moreira Gomes1
1Laboratório de Fisiologia e Bioquímica de Microorganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brasil
2Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brasil
3Departamento de Biofísica, Instituto de Física de São Carlos, Universidade de São Paulo, 60451-970, São Carlos, São Paulo, Brazil
4Laboratório de Melhoramento Genético Vegetal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brasil
5Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brasil

Tóm tắt

Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9‐kDa LTP from Capsicum annuum seeds that we call Ca‐LTP1. Ca‐LTP1 was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein's biological activity. Bidimensional electrophoresis of the 9‐kDa band, which corresponds to the purified Ca‐LTP1, showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of α‐helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca‐LTP1 from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca‐LTP1 indicated that it was mainly localized within dense vesicles. In addition, isolated Ca‐LTP1 exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca‐LTP1 also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca‐LTP1 is able to inhibit mammalian α‐amylase activity in vitro.

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Physiologia Plantarum

Tập 142 Số 3

233-246

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