MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells

JIANG Daifeng; et al

doi:10.13406/j.cnki.cyxb.2005.04.001

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MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells

更新时间：2024-01-17

- MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells
- Journal of Chongqing Medical University Issue 4, Pages: 485-488(2005)
- 作者机构：
  
  1. 重庆医科大学医学检验系实验诊断教研室
  2. 重庆医科大学基础医学研究所,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.04.001
  CLC： R735.7
- Published：2005，
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[1]蒋代凤,邱宗荫,李兴,黄开顺,滕永真,张晓萍.肝癌细胞线粒体蛋白的MALDI-TOF质谱鉴定方法研究[J].重庆医科大学学报,2005(04):485-488.

JIANG Daifeng, et al. MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells. [J]. Journal of Chongqing Medical University (4):485-488(2005)
[1]蒋代凤,邱宗荫,李兴,黄开顺,滕永真,张晓萍.肝癌细胞线粒体蛋白的MALDI-TOF质谱鉴定方法研究[J].重庆医科大学学报,2005(04):485-488. DOI： 10.13406/j.cnki.cyxb.2005.04.001.

JIANG Daifeng, et al. MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells. [J]. Journal of Chongqing Medical University (4):485-488(2005) DOI： 10.13406/j.cnki.cyxb.2005.04.001.

摘要

目的:探索胶上蛋白原位酶切、肽质量指纹图（PMF）分析以及数据库检索方法的优化条件

建立适合于亚细胞比较蛋白质组学研究的质谱鉴定策略。方法:从考马斯亮蓝染色的2-DE胶上取下牛血清白蛋白（BSA）标准蛋白进行胶内原位酶切

与基质（CHCA）混匀后进行MALDI-TOF MS分析;对来源于人肝细胞癌细胞株QGY-7703线粒体的差异表达的候选蛋白点L9

按照通过BSA标准蛋白优化的条件进行MALDI-TOF质谱鉴定;其肽质量指纹图（PMF）经MS-Fit检索。结果:经数据库检索

BSA的肽质量指纹图（PMF）匹配肽质量数为10/11

序列覆盖率为19%

说明实验条件完全可信。候选蛋白点L9的PMF数据经数据库检索后

排名前两位的蛋白均为OXCT（3-oxoacid CoA transferase 1 precursor）

其匹配肽段为9/13

序列覆盖率为24%;且OXCT的理论分子量（56kda）

理论等电点（7.1）

与胶上L9蛋白的位置相符。故候选蛋白点L9确认为OXCT（3-oxoacid CoA transferase 1 precursor）。结论:本文所确定的鉴定策略适用于线粒体比较蛋白质组学的研究。

Abstract

Objective:To explore the optimal conditions associated with proteolytic digestion in 2-DE gel

PMF analysis and database inquiry

and to construct a mass identification strategy in research for mitochondrial comparative proteomics.Methods:The BSA standard protein was incised from 2-DE gel by Coomassie Brilliant Blue dyeing and was mixed with matrix (CHCA)

then analyzed by MALDI-TOF MS.The differentially expressed mitochondria protein L9

an unknown protein obtained from hepatoma cells QGY-7703

was identified by condition with BSA optimizing.The PMF was inquired by MS-Fit database.Results:The BSA was identified in Masses Matched number 10/11 and Sequence Overcast 19% of PMF

which proved to the reliability of experimental conditions.By database Inquiry about PMF from L9 protein

OXCT(3-oxoacid CoA transferase 1 precursor)was identified with Masses Matched number 9/13 and Sequence Overcast 24%.The OXCT was in accord with position on gel.Conclusion:This method is applicable to research for mitochondrial comparative proteomics.

关键词

线粒体蛋白质组学肽质量指纹图（PMF）基质辅助激光解吸离子化-飞行时间质谱（MALDI-TOFMASS）二维电泳(2-DE

Keywords

mitochondriaproteomicsPeptide mass fingerprinting(PMF)Matrix assisted laser desorption ionization-time of flight mass spectroscopy(MALDI-TOF MS)Two-dimensional electrophoresis(2-DE)

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