Crystal structure of an antifungal osmotin-like protein from Calotropis procera and its effects on Fusarium solani spores, as revealed by atomic force microscopy: Insights into the mechanism of action

被引:34
作者
Ramos, Marcio V. [1 ]
de Oliveira, Raquel S. B. [1 ]
Pereira, Humberto M. [2 ,3 ]
Moreno, Frederico B. M. B. [3 ]
Lobo, Marina D. P.
Rebelo, Luciana M. [4 ]
Brandao-Neto, Jose [5 ]
de Sousa, Jeanlex S. [4 ]
Monteiro-Moreira, Ana C. O. [3 ]
Freitas, Cleverson D. T. [1 ]
Grangeiro, Thalles Barbosa [6 ]
机构
[1] Univ Fed Ceara, Ctr Ciencias, Dept Bioquim & Biol Mol, BR-60440900 Fortaleza, Ceara, Brazil
[2] Univ Sao Paulo, Inst Fis Sao Carlos, BR-13563120 Sao Paulo, Brazil
[3] Univ Fortaleza, Nucleo Biol Expt, Fortaleza, Ceara, Brazil
[4] Univ Fed Ceara, Ctr Ciencias, Dept Fis, BR-60440900 Fortaleza, Ceara, Brazil
[5] Diamond Light Source, Didcot OX11 0DE, Oxon, England
[6] Univ Fed Ceara, Ctr Ciencias, Dept Biol, BR-60440900 Fortaleza, Ceara, Brazil
关键词
Sodom apple; Calotropis procera; Apocynaceae; Protein; Thaumatin-like protein; THAUMATIN-LIKE PROTEIN; DISEASE RESISTANCE; XYLANASE INHIBITOR; RESOLUTION; IDENTIFICATION; MEMBRANE; GENES; CRYSTALLIZATION; OVEREXPRESSION; ELECTROSTATICS;
D O I
10.1016/j.phytochem.2015.09.012
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
CpOsm is an antifungal osmotin/thaumatin-like protein purified from the latex of Calotropis procera. The protein is relatively thermostable and retains its antifungal activity over a wide pH range; therefore, it may be useful in the development of new antifungal drugs or transgenic crops with enhanced resistance to phytopathogenic fungi. To gain further insight into the mechanism of action of CpOsm, its three-dimensional structure was determined, and the effects of the protein on Fusarium solani spores were investigated by atomic force microscopy (AFM). The atomic structure of CpOsm was solved at a resolution of 1.61 angstrom, and it contained 205 amino acid residues and 192 water molecules, with a final R-factor of 18.12% and an R-free of 21.59%. The CpOsm structure belongs to the thaumatin superfamily fold and is characterized by three domains stabilized by eight disulfide bonds and a prominent charged cleft, which runs the length of the front side of the molecule. Similarly to other antifungal thaumatin-like proteins, the cleft of CpOsm is predominantly acidic. AFM images of F. solani spores treated with CpOsm resulted in striking morphological changes being induced by the protein. Spores treated with CpOsm were wrinkled, and the volume of these cells was reduced by approximately 80%. Treated cells were covered by a shell of CpOsm molecules, and the leakage of cytoplasmic content from these cells was also observed. Based on the structural features of CpOsm and the effects that the protein produces on F. solani spores, a possible mechanism of action is suggested and discussed. (C) 2015 Elsevier Ltd. All rights reserved.
引用
收藏
页码:5 / 18
页数:14
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