论文部分内容阅读
通过对NCBI数据库中不同物种同源序列进行比对protein-protein BLAST,并利用计算机软件DiscoveryStudio3.1进行分子对接和分子动力学模拟,对天然人甲状旁腺激素(1 34)的3个氨基酸R25K26K27进行Q25E26L27置换,并对突变体的生物活性进行了评价。结果显示:突变后PTH(1 34)-(RKK-QEL)和突变前PTH(1 34)多肽主链叠合后均方根偏差RMSD值为2.509 3,说明两者主链结构构象差异不大;PTH(1 34)-(RKK-QEL)与受体蛋白PTH1R的相互作用能为599.253 kcal.mol 1,较天然PTH(1 34)的554.083 kcal.mol 1增强7.5%;氢键数目由32对增加至38对;PTH(1 34)-(RKK-QEL)能显著刺激UAMS-32P细胞RANKL基因的表达(P<0.01),并抑制OPG基因的表达(P<0.01);PTH(1 34)-(RKK-QEL)作用于UAMS-32P和小鼠原代股骨骨髓细胞共培养体系时,能显著刺激破骨细胞的形成(P<0.01),并且活性高于PTH(1 34)标准品。
By comparing the protein-protein BLAST of homologous sequences of different species in NCBI database and using the software of software DiscoveryStudio3.1 to molecular docking and molecular dynamics simulation, the three amino acids R25K26K27 of natural human parathyroid hormone (1 34) The Q25E26L27 substitution was performed and the biological activity of the mutant was evaluated. The results showed that the root mean square deviation (RMSD) of the PTH (1 34) - (RKK-QEL) and the pre-mutation PTH (1 34) peptide backbone after mutation was 2.509 3, indicating that there is no significant difference in the main structure between the two ; The interaction energy between PTH (1 34) - (RKK-QEL) and the receptor protein PTH1R was 599.253 kcal.mol 1, which was 7.5% higher than 554.083 kcal.mol 1 of native PTH (1 34); the number of hydrogen bonds was 32 PTH (1 34) - (RKK-QEL) significantly increased the expression of RANKL gene (P <0.01) and inhibited the expression of OPG gene in UAMS-32P cells (P <0.01) ) - (RKK-QEL) significantly stimulated the formation of osteoclasts (P <0.01) when UAMS-32P and mouse primary femur marrow coculture system were co-cultured, and its activity was higher than that of PTH .