人精浆小分子抗菌肽的分离鉴定
文献类型：期刊论文
| 作者 | 赵晖1,2; 申吉泓2; 李文辉3; 张云3; 李虹[*]1 |
| 刊名 | 四川大学学报( 医学版)
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| 出版日期 | 2008 |
| 卷号 | 39期号:5页码:842 - 846 |
| 关键词 | 精液 抗菌肽 分离纯化 精液凝固蛋白 |
| 通讯作者 | ydyyzh@263.net |
| 中文摘要 | 目的从人精浆中分离小分子抗菌肽,探讨精浆的抗菌机制。方法收集正常人精液,室温液化后10000r/min离心10min去除精子得到精浆。采用琼脂糖弥散法检测抗菌活性,采用对大肠杆菌(ATCC25922)的抗菌活性作为检测指标,通过阳离子交换柱SP-Sepharose分离,将有抗菌活性的洗脱产物冻干后用去离子水溶解,经过AKTA Superdex 75柱分离,收集小分子部分,应用反相高效液相色谱技术分离纯化多肽(RP-HPLC分离),检测各峰抗菌活性,基质辅助激光解吸质谱法检测相对分子质量,并鉴定其抗菌活性。结果从人精浆中初步分离出多个相对分子质量小于10×103的肽混合物,其中部分与精液凝固蛋白Ⅰ片段相对分子质量大小一致,命名为HSLAMs(Human semen low-molecular-mass antibacterial mixtures),具有抗大肠杆菌的活性。结论HSLAMs可能是人精浆天然免疫机制中重要的效应分子,精液凝固蛋白Ⅰ可能是其来源之一。 |
| 英文摘要 | Objective To isolate low-molecular-mass antibacterial mixtures from healthy human seminal plasma. Methods Semen was obtained by masturbation after at least three days of abstinence from healthy donors. Semen samples were allowed to liquefy at room temperature and then centrifuged at 10 000 r/min for 10 rain to separate spermatozoa from seminal plasma. High sensitive antimierobial activity was measured with radial diffusion assay. Antibacterial activity toward E. coli （ATCC25922） was monitored for each purification steps. The mixture of seminal plasma samples was applied to a SP-Sepharose column. Fractions which showed strong bactericidal activities, were combined and lyophilized. The lyophilized components were dissolved with Milli-Q water and applied to AKTA Superdex 75 column. Peak I of the Superdex 75 column, which showed antibacterial activity and represented the low-molecular-weight cationic fractions of the seminal plasma, was collected and lyophilized. Finally, peak I of the Superdex 75 column was applied to reverse phase HPLC C18 column. Fractions which showed strong antibacterial activity, were lyophilized and store at -20 ℃. The molecular weight of the low-molecule-mass antibacterial mixtures was determined by Matrix-assisted laser desorption/ionization （MALDI） mass spectrometry. Results The low-molecular-mass mixtures with obviously higher antibacterial activity, which were termed HSLAMs （Human semen low-molecular-mass antibacterial mixtures）, were isolated from the healthy human seminal plasma, Based on the mass spectrometry results, some molecules of RP-HPLC peaks were confirmed to be the semenogelin I derived peptides. Conclusion The low-molecule-mass antibacterial mixtures may play an important role in males innate immunity. Semenogelin I derived peptides may be one of the sources of the low-molecule-mass antimierobial mixtures in human seminal plasma. |
| 收录类别 | 其他 |
| 语种 | 中文 |
| 公开日期 | 2013-06-24 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7517]  |
| 专题 | 昆明动物研究所_动物活性蛋白多肽组学 昆明动物研究所_动物模型与人类重大疾病机理重点实验室 |
| 作者单位 | 1.四川大学华西医院泌尿外科,成都610041 2.昆明医学院第一附属医院泌尿外科 ,成都610041 3.中国科学院昆明动物研究所动物模型和人类疾病机理重点实验室 |
| 推荐引用方式 GB/T 7714 | 赵晖,申吉泓,李文辉,等. 人精浆小分子抗菌肽的分离鉴定[J]. 四川大学学报( 医学版),2008,39(5):842 - 846 . |
| APA | 赵晖,申吉泓,李文辉,张云,&李虹[*].(2008).人精浆小分子抗菌肽的分离鉴定.四川大学学报( 医学版),39(5),842 - 846 . |
| MLA | 赵晖,et al."人精浆小分子抗菌肽的分离鉴定".四川大学学报( 医学版) 39.5(2008):842 - 846 . |
入库方式： OAI收割
来源：昆明动物研究所
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