摘要
目的合成肺鳞癌靶点蛋白基质金属蛋白酶9(MMP9)的靶向多肽并进行验证,旨在研发一种新型MMP9靶向多肽。方法使用Molecular Operating Environment软件将蛋白质结构数据库(PDB)中获得的肺鳞癌靶点蛋白MMP9及其配体晶体结构进行优化,通过分子对接确定MMP9蛋白及其配体的相互作用位点,并结合London dG打分函数(S score)确认关键残基为Tyr49、Ser50和Asn55;选取模板多肽后进行突变替换,进行蛋白-蛋白对接后,选取S score最低的6条候选多肽。进行合成可行性评估后,筛选出3条候选多肽,与模板多肽一同进行合成。采用高效液相色谱法对合成的4条多肽进行纯度及结构鉴定。取人肺鳞癌细胞NCI-H226分成SP-模板多肽组(加入SP-模板多肽)、SP-1多肽组(加入SP-1多肽)、SP-2多肽组(加入SP-2多肽)、SP-3多肽组(加入SP-3多肽)、对照组(加入MMP9抗体和荧光肽),采集荧光图像后,通过Image J软件分析各组荧光强度,评估不同候选多肽和人肺鳞癌细胞NCI-H226的亲和性。结果成功合成4条目标多肽,质谱验证其结构正确,纯度均高于95%。SP-模板多肽组、SP-1多肽组、SP-2多肽组、SP-3多肽组、对照组荧光强度分别为1.48±0.15、1.14±0.12、1.85±0.07、0.68±0.20、1.00±0.13,其中SP-2多肽组荧光强度均高于其余各组(P均<0.05)。结论成功合成了一种新型MMP9靶向多肽SP-2,其结构正确、纯度较高,且与人肺鳞癌细胞亲和性较高。
Objective To synthesize and verify a matrix metalloproteinase-9(MMP9)-targeting peptide for lung squamous cell carcinoma,aiming to develop a novel MMP9-targeting peptide.Methods The crystal structures of MMP9 and its ligand were obtained from the Protein Data Bank(PDB)and optimized using Molecular Operating Environment soft⁃ware.Molecular docking was performed to identify interaction sites between MMP9 and its ligand.Key residues(Tyr49,Ser50,and Asn55)were confirmed using the London dG scoring function(S score).A template peptide was selected and mutated,followed by protein-protein docking to screen six candidate peptides with the lowest S scores.After synthetic fea⁃sibility assessment,three candidate peptides were selected for synthesis along with the template peptide.The purity and structure of the four synthesized peptides were characterized by high-performance liquid chromatography(HPLC).Human lung squamous cell carcinoma cells(NCI-H226)were divided into five groups:SP-template peptide group,SP-1 peptide group,SP-2 peptide group,SP-3 peptide group,and control group(treated with MMP9 antibody and fluorescent pep⁃tide).Fluorescence images were acquired,and fluorescence intensity was analyzed using ImageJ software to evaluate the affinity of the candidate peptides for NCI-H226 cells.Results Four target peptides were successfully synthesized,with their structures validated by mass spectrometry and purity levels exceeding 95%.The fluorescence intensities of the SPtemplate peptide group,SP-1 peptide group,SP-2 peptide group,SP-3 peptide group,and control group were 1.48±0.15,1.14±0.12,1.85±0.07,0.68±0.20,and 1.00±0.13,respectively,with the SP-2 peptide group exhibiting significantly higher fluorescence intensity than all other groups(all P<0.05).Conclusions The novel MMP9-targeting peptide SP-2 was successfully synthesized.The synthesized peptide was verified to have the correct structure and high puri⁃ty,and it demonstrated high binding affinity toward human lung squamous cell carcinoma cells.
作者
臧国杰
霍李娜
武美元
刘巾玮
ZANG Guojie;HUO Lina;WU Meiyuan;LIU Jinwei(School of Pharmacy,Inner Mongolia Medical University,Hohhot 010107,China;不详)
出处
《山东医药》
2026年第1期49-53,共5页
Shandong Medical Journal
基金
内蒙古自治区卫生健康委员会公立医院科研联合基金科技项目(2023GLLHO300)
内蒙古医科大学联合项目(YKD2023LH006)。
关键词
基质金属蛋白酶9
肺鳞癌
靶向多肽
分子对接
固相合成
matrix metalloproteinase 9
lung squamous cell carcinoma
targeted peptides
molecular docking
solid-phase synthesis
