Citation: ZHOU Yuan, ZHANG Shen, YUAN Huiming, ZHANG Lihua, ZHANG Yukui. Establishment and application of integrated platform for proteome quantification[J]. Chinese Journal of Chromatography, ;2014, 32(4): 355-360. doi: 10.3724/SP.J.1123.2014.01002 shu

Establishment and application of integrated platform for proteome quantification

ZHOU Yuan ^1,2 ,
ZHANG Shen ^1,2 ,
YUAN Huiming ¹ ,
ZHANG Lihua ^1,, ,
ZHANG Yukui ¹

1.
1. Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences; National Chromatographic Research and Analysis Center, Dalian 116023, China;

Corresponding author: ZHANG Lihua,
Received Date: 2 January 2014
Available Online: 3 March 2014
Fund Project: 国家重大科学研究计划(2012CB910604)；国家重大科学仪器设备开发专项(2012YQ120044-2)；国家自然科学基金项目(21190043). (2012CB910604)；国家重大科学仪器设备开发专项(2012YQ120044-2)；国家自然科学基金项目(21190043)

To improve the accuracy, throughput and automation of proteome quantification analysis, an integrated platform including a microflow mixed-bed ion exchange column, a hydrophilic immobilized enzymatic reactor (hIMER) and nanoRPLC-electrospray ionization (ESI)-MS/MS system was established. Online separation and digestion of dimethylated proteins, followed by peptide separation, identification and quantification can be realized automatically by this platform. High and light dimethyl-labeled (H/L) proteins with the mass ratio of 1:1 were used to evaluate the quantification performance of the platform. The results showed that the dimethyl labeling efficiency at protein level was 90%. The incomplete digestion resulting from 10 min online digestion by the hIMER column and the non-specific adsorption of protein digests on the column had little adverse effect on the accuracy of protein quantification results. The mean value of H/L (mass ratio) of all the quantified proteins was 1.01. This platform was finally applied to analyze the different protein expression levels of two mice hepatocarcinoma ascites cell lines with high and low lymph node metastasis rates (Hca-F and Hca-P cell lines). Finally 15 up-regulated and 12 down-regulated proteins (Hca-F/Hca-P) were successfully obtained. All these results demonstrated that the integrated platform can be used for proteome quantification with the advantages of high accuracy and high throughput.
1. [1]
  [1] Yates J R Ⅲ, Washburn M P. Anal Chem, 2013, 85(19): 8881
2. [2]
  [2] Merrill A E, Coon J J. Curr Opin Chem Biol, 2013, 17(5): 779
3. [3]
  [3] Boersema P J, Raijmakers R, Lemeer S, et al. Nat Protoc, 2009, 4(4): 484
4. [4]
  [4] Raijmakers R, Berkers C R, de Jong A, et al. Mol Cell Proteomics, 2008, 7(9): 1755
5. [5]
  [5] Wang F J, Chen R, Zhu J, et al. Anal Chem, 2010, 82(7): 3007
6. [6]
  [6] Ong S E, Mann M. Nat Chem Biol, 2005, 1(5): 252
7. [7]
  [7] Bantscheff M, Schirle M, Sweetman G, et al. Anal Bioanal Chem, 2007, 389(4): 1017
8. [8]
  [8] Zhou Y, Shan Y C, Zhang L H, et al. Chinese Journal of Chromatography (周愿, 单亦初, 张丽华, 等. 色谱), 2013, 31(6): 496
9. [9]
  [9] Tian R J, Wang S, Elisma F, et al. Mol Cell Proteomics, 2011, 10(2), DOI: 10.1074/mcp. M110.000679
10. [10]
  [10] Yuan H M, Zhou Y, Zhang L H, et al. J Chromatogr A, 2009, 1216(44): 7478
11. [11]
  [11] Yuan H M, Zhang L H, Hou C Y, et al. Anal Chem, 2009, 81(21): 8708
12. [12]
  [12] Yuan H M, Zhou Y, Xia S M, et al. Anal Chem, 2012, 84(11): 5124
13. [13]
  [13] Xia S M, Tao D Y, Yuan H M, et al. J Sep Sci, 2012, 35(14): 1764
14. [14]
  [14] Jiang H, Yuan H M, Liang Y, et al. J Chromatogr A, 2012, 1246: 111
15. [15]
  [15] Wang F J, Wei X L, Zhou H, et al. Proteomics, 2012, 12(21): 3129
16. [16]
  [16] Sun M Z, Liu S Q, Tang J W, et al. Proteomics, 2009, 9(12): 3285
17. [17]
  [17] Zhou Y, Shan Y C, Wu Q, et al. Anal Chem, 2013, 85(22): 10658
18. [18]
  [18] Liu W, Ma Y, Huang L, et al. Mol Biol Rep, 2010, 37(7): 3207
19. [19]
  [19] Wu N, Liu S Q, Guo C M, et al. Clin Transl Oncol, 2013, 15(2): 106
1. [1]
  Xinyi Hong , Tailing Xue , Zhou Xu , Enrong Xie , Mingkai Wu , Qingqing Wang , Lina Wu . Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment. University Chemistry, 2024, 39(4): 351-360. doi: 10.3866/PKU.DXHX202310010
2. [2]
  Ling Bai , Limin Lu , Xiaoqiang Wang , Dongping Wu , Yansha Gao . Exploration and Practice of Teaching Reforms in “Quantitative Analytical Chemistry” under the Perspective of New Agricultural Science. University Chemistry, 2024, 39(3): 158-166. doi: 10.3866/PKU.DXHX202308101
3. [3]
  Yongjie ZHANG , Bintong HUANG , Yueming ZHAI . Research progress of formation mechanism and characterization techniques of protein corona on the surface of nanoparticles. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2318-2334. doi: 10.11862/CJIC.20240247
4. [4]
  Zeyu XU , Anlei DANG , Bihua DENG , Xiaoxin ZUO , Yu LU , Ping YANG , Wenzhu YIN . Evaluation of the efficacy of graphene oxide quantum dots as an ovalbumin delivery platform and adjuvant for immune enhancement. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1065-1078. doi: 10.11862/CJIC.20240099
5. [5]
  Min Gu , Huiwen Xiong , Liling Liu , Jilie Kong , Xueen Fang . Rapid Quantitative Detection of Procalcitonin by Microfluidics: An Instrumental Analytical Chemistry Experiment. University Chemistry, 2024, 39(4): 87-93. doi: 10.3866/PKU.DXHX202310120
6. [6]
  Wentao Lin , Wenfeng Wang , Yaofeng Yuan , Chunfa Xu . Concerted Nucleophilic Aromatic Substitution Reactions. University Chemistry, 2024, 39(6): 226-230. doi: 10.3866/PKU.DXHX202310095
7. [7]
  Hongwei Ma , Fang Zhang , Hui Ai , Niu Zhang , Shaochun Peng , Hui Li . Integrated Crystallographic Teaching with X-ray,TEM and STM. University Chemistry, 2024, 39(3): 5-17. doi: 10.3866/PKU.DXHX202308107
8. [8]
  Jia Huo , Jia Li , Yongjun Li , Yuzhi Wang . Ideological and Political Design of Physical Chemistry Teaching: Chemical Potential of Any Component in an Ideal-Dilute Solution. University Chemistry, 2024, 39(2): 14-20. doi: 10.3866/PKU.DXHX202307075
9. [9]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
10. [10]
  Zhenming Xu , Yibo Wang , Zhenhui Liu , Duo Chen , Mingbo Zheng , Laifa Shen . Experimental Design of Computational Materials Science and Computational Chemistry Courses Based on the Bohrium Scientific Computing Cloud Platform. University Chemistry, 2025, 40(3): 36-41. doi: 10.12461/PKU.DXHX202403096
11. [11]
  Zhonghua Xi , Xuanfeng Kong , Jinyue Yang , Bin Liu , Tingyu Zhu , Hui Zhang , Wenwei Zhang . Construction of Public Teaching Instrument Platform and Exploration of Opening Mechanism. University Chemistry, 2024, 39(7): 200-206. doi: 10.12461/PKU.DXHX202405123
12. [12]
  Siming Bian , Sijie Luo , Junjie Ou . Application of van Deemter Equation in Instrumental Analysis Teaching: A New Type of Core-Shell Stationary Phase. University Chemistry, 2025, 40(3): 381-386. doi: 10.12461/PKU.DXHX202406087
13. [13]
  Yue Zhao , Yanfei Li , Tao Xiong . Copper Hydride-Catalyzed Nucleophilic Additions of Unsaturated Hydrocarbons to Aldehydes and Ketones. University Chemistry, 2024, 39(4): 280-285. doi: 10.3866/PKU.DXHX202309001
14. [14]
  Yuqiao Zhou , Weidi Cao , Shunxi Dong , Lili Lin , Xiaohua Liu . Study on the Teaching Reformation of Practical X-ray Crystallography. University Chemistry, 2024, 39(3): 23-28. doi: 10.3866/PKU.DXHX202303003
15. [15]
  Xingchao Zhao , Xiaoming Li , Ming Liu , Zijin Zhao , Kaixuan Yang , Pengtian Liu , Haolan Zhang , Jintai Li , Xiaoling Ma , Qi Yao , Yanming Sun , Fujun Zhang . 倍增型全聚合物光电探测器及其在光电容积描记传感器上的应用. Acta Physico-Chimica Sinica, 2025, 41(1): 2311021-. doi: 10.3866/PKU.WHXB202311021
16. [16]
  Shiyan Cheng , Yonghong Ruan , Lei Gong , Yumei Lin . Research Advances in Friedel-Crafts Alkylation Reaction. University Chemistry, 2024, 39(10): 408-415. doi: 10.12461/PKU.DXHX202403024
17. [17]
  Weina Wang , Lixia Feng , Fengyi Liu , Wenliang Wang . Computational Chemistry Experiments in Facilitating the Study of Organic Reaction Mechanism: A Case Study of Electrophilic Addition of HCl to Asymmetric Alkenes. University Chemistry, 2025, 40(3): 206-214. doi: 10.12461/PKU.DXHX202407022
18. [18]
  Fan Wu , Wenchang Tian , Jin Liu , Qiuting Zhang , YanHui Zhong , Zian Lin . Core-Shell Structured Covalent Organic Framework-Coated Silica Microspheres as Mixed-Mode Stationary Phase for High Performance Liquid Chromatography. University Chemistry, 2024, 39(11): 319-326. doi: 10.12461/PKU.DXHX202403031
19. [19]
  Qiying Xia , Guokui Liu , Yunzhi Li , Yaoyao Wei , Xia Leng , Guangli Zhou , Aixiang Wang , Congcong Mi , Dengxue Ma . Construction and Practice of “Teaching-Learning-Assessment Integration” Model Based on Outcome Orientation: Taking “Structural Chemistry” as an Example. University Chemistry, 2024, 39(10): 361-368. doi: 10.3866/PKU.DXHX202311007
20. [20]
  Ji-Quan Liu , Huilin Guo , Ying Yang , Xiaohui Guo . Calculation and Discussion of Electrode Potentials in Redox Reactions of Water. University Chemistry, 2024, 39(8): 351-358. doi: 10.3866/PKU.DXHX202401031

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(188)
HTML views(20)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142