CREA Research Centre for Vegetable and Ornamental Crops, Pontecagnano Faiano, Italy.
Institute of Biostructures and Bioimaging, CNR, Naples, Italy.
Sci Rep. 2019 Feb 7;9(1):1607. doi: 10.1038/s41598-018-38301-z.
Non-specific lipid transfer proteins (nsLTPs) are characterized by an eight-cysteine motif backbone that is stabilized by four disulphide bonds. The strong interest towards this protein family is mainly due to the fact that nsLTPs are involved in many biological processes and have been identified as major human allergens. Since tomato (Solanum lycopersicum L.) is one of the most consumed and allergenic vegetables, a full characterization of this family is needed. In this study, hidden Markov model profiles were used to identify nsLTPs within the tomato protein complement. Following manual curation, 64 nsLTP genes were classified into six sub-families. Furthermore, nsLTP gene structure, distribution and arrangement along tomato chromosomes were investigated. Available RNA-seq expression profile data and Real-Time PCR analyses were used to derive expression patterns of tomato nsLTPs in different tissues/organs. Non-specific LTP genes with high level of expression in tomato fruits were filtered out since they could play a key role in tomato allergenicity. Among these genes was Solyc10g075090 that encodes the allergen Sola l 3. Finally, cloning, heterologous expression, purification and biochemical characterization of the recombinant protein Sola l 3 was performed.
非特异性脂质转移蛋白(nsLTPs)的特征是具有由四个二硫键稳定的八个半胱氨酸基序骨干。人们对这种蛋白质家族的强烈兴趣主要是因为 nsLTPs 参与了许多生物过程,并且已被确定为主要的人类过敏原。由于番茄(Solanum lycopersicum L.)是最常食用和最易引起过敏的蔬菜之一,因此需要对该家族进行全面表征。在这项研究中,隐马尔可夫模型图谱被用于鉴定番茄蛋白质组中的 nsLTPs。经过手动整理,将 64 个 nsLTP 基因分为六个亚家族。此外,还研究了 nsLTP 基因结构、分布和在番茄染色体上的排列。利用可用的 RNA-seq 表达谱数据和实时 PCR 分析,得出了番茄 nsLTPs 在不同组织/器官中的表达模式。从番茄果实中高度表达的非特异性 LTP 基因中筛选出来,因为它们可能在番茄过敏中发挥关键作用。其中一个基因是 Solyc10g075090,它编码过敏原 Sola l 3。最后,对重组蛋白 Sola l 3 进行了克隆、异源表达、纯化和生化特性分析。