背景:癌症是全球范围内的主要死亡原因和重大公共卫生问题。标准治疗方法,如化疗,放射治疗,手术有时才有效.因此,癌症治疗需要新的治疗方法。海葵放线孔蛋白是具有膜分解活性的成孔毒素(PFTs)。RTX-A是一种与膜磷脂相互作用的PFT,导致孔隙形成。以分泌形式合成重组蛋白有几个优点,包括蛋白质溶解性和易于纯化。在这项研究中,我们旨在发现合适的信号肽,以分泌形式在枯草芽孢杆菌中产生RTX-A。
方法:从信号肽网络服务器中选择信号肽。使用SignalP服务器评估所选信号肽的概率和分泌途径。ProtParam和Protein-sol用于预测物理化学性质和溶解度。AlgPred用于预测与合适的信号肽连接的RTX-A的变应原性。非过敏性,稳定,并选择与蛋白质融合的可溶性信号肽,使用GORIV和I-TASSER预测了它们的二级和三级结构,分别。PROCHECK服务器执行3D结构的验证。
结果:根据生物信息学分析,与RTX-A连接的OSMY_ECOLI和MAE_ECOLI的融合形式被鉴定为合适的信号肽。带有信号肽的最终蛋白质是稳定的,可溶性,对人体不过敏。此外,他们有适当的二级和三级结构。
结论:上述肽的信号对于合理化分泌型和可溶性RTX-A似乎是理想的。因此,本研究中发现的信号肽需要通过实验研究和专利进一步研究。
BACKGROUND: Cancer is a leading cause of death and a significant public health issue worldwide. Standard treatment methods such as chemotherapy, radiotherapy, and surgery are only sometimes effective. Therefore, new therapeutic approaches are needed for cancer treatment. Sea anemone actinoporins are pore-forming toxins (PFTs) with membranolytic activities. RTX-A is a type of PFT that interacts with membrane phospholipids, resulting in pore formation. The synthesis of recombinant proteins in a secretory form has several advantages, including protein solubility and easy purification. In this study, we aimed to discover suitable signal peptides for producing RTX-A in Bacillus subtilis in a secretory form.
METHODS: Signal peptides were selected from the Signal Peptide Web Server. The probability and secretion pathways of the selected signal peptides were evaluated using the SignalP server. ProtParam and Protein-sol were used to predict the physico-chemical properties and solubility. AlgPred was used to predict the allergenicity of RTX-A linked to suitable signal peptides. Non-allergenic, stable, and soluble signal peptides fused to proteins were chosen, and their secondary and tertiary structures were predicted using GOR IV and I-TASSER, respectively. The PROCHECK server performed the validation of 3D structures.
RESULTS: According to bioinformatics analysis, the fusion forms of OSMY_ECOLI and MALE_ECOLI linked to RTX-A were identified as suitable signal peptides. The final proteins with signal peptides were stable, soluble, and non-allergenic for the human body. Moreover, they had appropriate secondary and tertiary structures.
CONCLUSIONS: The signal above peptides appears ideal for rationalizing secretory and soluble RTX-A. Therefore, the signal peptides found in this study should be further investigated through experimental researches and patents.