纳升级反相液相色谱-串联质谱法分析蜈蚣提取蛋白质
结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱（nanoRPLC-MS/MS）法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质。建立的nanoLC分析条件为：上样8μL,经Chrom XP-C18毛细管柱（350μm×0.5 mm,3μm）,2%乙腈（0.1%甲酸）-98%乙腈（0.1%甲酸）梯度洗脱90 min;采用ESI-Q-TOF MS,并在IDA采集模式下分析鉴定蜈蚣提取蛋白质。胶内酶解鉴定到72种蛋白质,直接酶解鉴定到97种蛋白质,二者含26种相同的蛋白质。通过数据库比对,分析了蜈蚣提取蛋白质的生物进程、细胞组分以及功能活性。得到蜈蚣蛋白质具有结合、...
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| Published in | 分析化学 Vol. 42; no. 2; pp. 239 - 243 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
南京中医药大学药学院,南京,210023%南京中医药大学中医学一级学科,南京,210023
2014
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0253-3820 |
| DOI | 10.3724/SP.J.1096.2014.30894 |
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| Abstract | 结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱（nanoRPLC-MS/MS）法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质。建立的nanoLC分析条件为：上样8μL,经Chrom XP-C18毛细管柱（350μm×0.5 mm,3μm）,2%乙腈（0.1%甲酸）-98%乙腈（0.1%甲酸）梯度洗脱90 min;采用ESI-Q-TOF MS,并在IDA采集模式下分析鉴定蜈蚣提取蛋白质。胶内酶解鉴定到72种蛋白质,直接酶解鉴定到97种蛋白质,二者含26种相同的蛋白质。通过数据库比对,分析了蜈蚣提取蛋白质的生物进程、细胞组分以及功能活性。得到蜈蚣蛋白质具有结合、酶催化、肌动等功能活性,其中结合活性的蛋白质占的比例较大,且多为能量代谢进程中的相关酶类。 |
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| AbstractList | 结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱（nanoRPLC-MS/MS）法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质。建立的nanoLC分析条件为：上样8μL,经Chrom XP-C18毛细管柱（350μm×0.5 mm,3μm）,2%乙腈（0.1%甲酸）-98%乙腈（0.1%甲酸）梯度洗脱90 min;采用ESI-Q-TOF MS,并在IDA采集模式下分析鉴定蜈蚣提取蛋白质。胶内酶解鉴定到72种蛋白质,直接酶解鉴定到97种蛋白质,二者含26种相同的蛋白质。通过数据库比对,分析了蜈蚣提取蛋白质的生物进程、细胞组分以及功能活性。得到蜈蚣蛋白质具有结合、酶催化、肌动等功能活性,其中结合活性的蛋白质占的比例较大,且多为能量代谢进程中的相关酶类。 结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱(nanoRPLC-MS/MS)法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质.建立的nanoLC分析条件为:上样8μL,经Chrom XP-C18毛细管柱(350 μm×0.5 mm,3μm),2％乙腈(0.1％甲酸)-98％乙腈(0.1％甲酸)梯度洗脱90 min;采用ESI-Q-TOF MS,并在IDA采集模式下分析鉴定蜈蚣提取蛋白质.胶内酶解鉴定到72种蛋白质,直接酶解鉴定到97种蛋白质,二者含26种相同的蛋白质.通过数据库比对,分析了蜈蚣提取蛋白质的生物进程、细胞组分以及功能活性.得到蜈蚣蛋白质具有结合、酶催化、肌动等功能活性,其中结合活性的蛋白质占的比例较大,且多为能量代谢进程中的相关酶类. |
| Author | 陈霞 文红梅 刘睿 朱栋 李伟 周红光 吴勉华 |
| AuthorAffiliation | 南京中医药大学药学院,南京210023 南京中医药大学中医学一级学科,南京210023 |
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| Author_FL | WEN Hong-Mei ZHU Dong CHEN Xia LIU Rui WU Mian-Hua LI Wei ZHOU Hong-Guang |
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| DOI | 10.3724/SP.J.1096.2014.30894 |
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| Keywords | Proteins Scolopendra Polyacrylamide gel electrophoresis 纳升级反相液相色谱-串联质谱 聚丙烯酰胺凝胶电泳 蜈蚣 Nanoflow reversed phase liquid chromatography-tandem mass spectrometry 蛋白质 |
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| Notes | Nanoflow reversed phase liquid chromatography-tandem mass spectrometry; Scolopendra; Proteins; Polyacrylamide gel electrophoresis CHEN Xia, WEN Hong-Mei, LIU Rui, ZHU Dong, LI Wei, ZHOU Hong-Guang, WU Mian-Hua 1 （ School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China） 2（ Traditional Chinese Medicine Department of Nanjing University of Chinese Medicine, Nanjing 210023, China） 22-1125/O6 Anovel nanoLC-MS/MS method coupled with SDS-PAGE was developed for analyzing extracted proteins of gel-based and gel-free digestion products from Scolopendra. NanoLC separation was performed on aChrom XP-C18 capillary column （350μm×0.5 mm,3μm）using a gradient elution program of 2% acetonitrile solution （containing 0. 1% formic acid） and 98% acetonitrile solution （containing 0.1% formic acid） as the mobile phase with a flow rate of 300 nL/min and theinjection volume of 8 μL. Quadruple-time-of- flight mass spectrometer （Q-TOF-MS） and electrospray ion source （ESI） were applied for qualitative analysis unde |
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| Publisher | 南京中医药大学药学院,南京,210023%南京中医药大学中医学一级学科,南京,210023 |
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| Snippet | 结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱（nanoRPLC-MS/MS）法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质。建立的nanoLC分析条件为：上样8μL,经Chrom XP-C18毛细管柱（350μm×0.5... 结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱(nanoRPLC-MS/MS)法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质.建立的nanoLC分析条件为:上样8μL,经Chrom XP-C18毛细管柱(350 μm×0.5... |
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| SubjectTerms | 纳升级反相液相色谱-串联质谱 聚丙烯酰胺凝胶电泳 蛋白质 蜈蚣 |
| Title | 纳升级反相液相色谱-串联质谱法分析蜈蚣提取蛋白质 |
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