运用反相高效液相色谱(RP-HPLC)对酶解螺蛳腹足肌得到的血管紧张素转换酶(ACE)抑制肽进行两步分离提纯,第一步主要得到8个组分;选取其中活性最高的组分进一步分离,得到2个组分,其中活性较高组分的ACE半抑制浓度为43.5μm o l/L,基本为...运用反相高效液相色谱(RP-HPLC)对酶解螺蛳腹足肌得到的血管紧张素转换酶(ACE)抑制肽进行两步分离提纯,第一步主要得到8个组分;选取其中活性最高的组分进一步分离,得到2个组分,其中活性较高组分的ACE半抑制浓度为43.5μm o l/L,基本为单一肽组分。对提纯的组分分别使用高效液相色谱/电喷雾离子质谱法(HPLC/E S I-M S)和基质辅助激光解吸电离飞行时间质谱法(MALD I-TO F M S)进行分析,同时结合氨基酸组成分析结果,最终得到的肽链一级结构为Lys-G lu-Ile-Trp(KE IW),符合已知的高活性ACE抑制肽的结构规律。经过对两种方法分析过程的比较,认为E S I-M S可以得到多方面的信息,但无法确定肽的序列;MALD I-TO F M S可以得到精确的二级质谱图(m/z精确至0.000 1),从而可以得到确定的肽的序列。展开更多
The progress of science is in part attributed to the development of new methodologies. The application of mass spectrometry (MS) and nuclear magnetic resonance (NMR) in the study of biological macromolecules is such a...The progress of science is in part attributed to the development of new methodologies. The application of mass spectrometry (MS) and nuclear magnetic resonance (NMR) in the study of biological macromolecules is such a representative example, which is the subject of this year’s Nobel prize award in chemistry shared by three scientists. The works include the development of soft desorption ionization methods for mass spectrometric analysis of biological macromolecules and the development of NMR for determining the three-dimensional structure of biological macromolecules in solution. These developments revolutionized the analytical methods for biomolecules such as proteins and facilitate the study of biological macromolecules so much enough to have deep effects on the whole life sciences.展开更多
文摘运用反相高效液相色谱(RP-HPLC)对酶解螺蛳腹足肌得到的血管紧张素转换酶(ACE)抑制肽进行两步分离提纯,第一步主要得到8个组分;选取其中活性最高的组分进一步分离,得到2个组分,其中活性较高组分的ACE半抑制浓度为43.5μm o l/L,基本为单一肽组分。对提纯的组分分别使用高效液相色谱/电喷雾离子质谱法(HPLC/E S I-M S)和基质辅助激光解吸电离飞行时间质谱法(MALD I-TO F M S)进行分析,同时结合氨基酸组成分析结果,最终得到的肽链一级结构为Lys-G lu-Ile-Trp(KE IW),符合已知的高活性ACE抑制肽的结构规律。经过对两种方法分析过程的比较,认为E S I-M S可以得到多方面的信息,但无法确定肽的序列;MALD I-TO F M S可以得到精确的二级质谱图(m/z精确至0.000 1),从而可以得到确定的肽的序列。
文摘目的:通过解吸电喷雾电离质谱成像(desorption electrospray ionization-mass spectrometry imaging,DESI-MSI)研究口腔鳞状细胞癌(oral squamous cell carcinomas,OSCC)的潜在生物标志物和可能的代谢途径。方法:本研究纳入的10例OSCC病例均为在江苏大学附属医院口腔颌面外科就诊住院且接受手术治疗的原发病例。收集组织样本用于DESI-MSI检测,进一步采用多元及单变量统计分析筛选差异代谢物,然后用受试者工作特征曲线(receiver operating characteristic curve,ROC曲线)分析差异代谢物的诊断能力。最后,进行京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢通路分析确定相关的代谢通路。结果:OSCC肿瘤组织与正常组织共鉴定出144种差异代谢物,大多数为脂肪酸。ROC曲线分析发现这144种代谢物曲线下的面积(area under the curve,AUC)值均大于0.7。KEGG代谢通路分析发现,与OSCC发生相关的通路为不饱和脂肪酸的生物合成以及脂肪酸的生物合成。结论:基于DESI-MSI筛选了具有诊断价值的差异代谢物,发现OSCC组织存在不饱和脂肪酸的生物合成以及脂肪酸的生物合成代谢紊乱,可能与OSCC的发生发展密切相关。
文摘The progress of science is in part attributed to the development of new methodologies. The application of mass spectrometry (MS) and nuclear magnetic resonance (NMR) in the study of biological macromolecules is such a representative example, which is the subject of this year’s Nobel prize award in chemistry shared by three scientists. The works include the development of soft desorption ionization methods for mass spectrometric analysis of biological macromolecules and the development of NMR for determining the three-dimensional structure of biological macromolecules in solution. These developments revolutionized the analytical methods for biomolecules such as proteins and facilitate the study of biological macromolecules so much enough to have deep effects on the whole life sciences.
