Hamman-Khalifa Abdelmounim, Castro Antonio Jesús, Jiménez-López José Carlos, Rodríguez-García María Isabel, Alché Juan de Dios
Department of Biochemistry, Cell and Molecular Biology of Plants, Estación, Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), Profesor Albareda 1, 18008, Granada, Spain.
BMC Plant Biol. 2008 Jan 25;8:10. doi: 10.1186/1471-2229-8-10.
Pollens from different olive (Olea europaea L.) cultivars have been shown to differ significantly in their content in Ole e 1 and in their overall allergenicity. This allergen is, in addition, characterized by a high degree of polymorphism in its sequence. The purpose of this study is to evaluate the putative presence of divergences in Ole e 1 sequences from different olive cultivars.
RNA from pollen individually collected from 10 olive cultivars was used to amplify Ole e 1 sequences by RT-PCR, and the sequences were analyzed by using different bioinformatics tools. Numerous nucleotide substitutions were detected throughout the sequences, many of which resulted in amino acid substitutions in the deduced protein sequences. In most cases variability within a single variety was much lower than among varieties. Key amino acid changes in comparison with "canonical" sequences previously described in the literature included: a) the substitution of C19-relevant to the disulphide bond structure of the protein-, b) the presence of an additional N-glycosylation motif, and c) point substitutions affecting regions of Ole e 1 already described like relevant for the immunogenicity/allergenicity of the protein.
Varietal origin of olive pollen is a major factor determining the diversity of Ole e 1 variants. We consider this information of capital importance for the optimal design of efficient and safe allergen formulations, and useful for the genetic engineering of modified forms of the allergen among other applications.
已表明来自不同油橄榄(油橄榄属)品种的花粉在其油橄榄蛋白1(Ole e 1)含量及其整体致敏性方面存在显著差异。此外,这种过敏原的序列具有高度多态性。本研究的目的是评估不同油橄榄品种的Ole e 1序列中是否存在差异。
从10个油橄榄品种单独收集的花粉中的RNA用于通过逆转录聚合酶链反应(RT-PCR)扩增Ole e 1序列,并使用不同的生物信息学工具对序列进行分析。在整个序列中检测到许多核苷酸替换,其中许多导致推导的蛋白质序列中的氨基酸替换。在大多数情况下,单个品种内的变异性远低于品种间的变异性。与文献中先前描述的“典型”序列相比,关键的氨基酸变化包括:a)与蛋白质二硫键结构相关的C19的替换,b)存在额外的N-糖基化基序,以及c)影响已描述的与蛋白质免疫原性/致敏性相关的Ole e 1区域的点替换。
油橄榄花粉的品种来源是决定Ole e 1变体多样性的主要因素。我们认为这些信息对于高效和安全的过敏原制剂的优化设计至关重要,并且在其他应用中对于过敏原修饰形式的基因工程也很有用。