Citation: LI Nannan, ZHOU Lianqi, MAO Xinli, ZHANG Jiao, WEI Junying, LIN Hongjun, LI Jiabin, TIAN Fang, ZHANG Yangjun, QIAN Xiaohong. A novel method for absolute protein quantification using ¹⁸O isotope labeled concatamers of Q peptides combined with isotope dilution-multiple reaction monitoring mass spectrometry[J]. Chinese Journal of Chromatography, ;2013, 31(6): 522-530. doi: 10.3724/SP.J.1123.2013.03057 shu

A novel method for absolute protein quantification using ¹⁸O isotope labeled concatamers of Q peptides combined with isotope dilution-multiple reaction monitoring mass spectrometry

LI Nannan ^1,2 ,
ZHOU Lianqi ² ,
MAO Xinli ² ,
ZHANG Jiao ² ,
WEI Junying ² ,
LIN Hongjun ² ,
LI Jiabin ² ,
TIAN Fang ² ,
ZHANG Yangjun ^2,, ,
QIAN Xiaohong ²

1.
1. Guangxi Medical University, Nanning 530021, China;

Corresponding author: ZHANG Yangjun,
Received Date: 31 March 2013
Available Online: 3 May 2013
Fund Project: 国家重大科学计划项目(2012CB910603,2010CB912704) (2012CB910603,2010CB912704)国家重大科学仪器设备开发专项项目(2011YQ030139,2011YQ06008408,2011YQ09000504,2012YQ12004407) (2011YQ030139,2011YQ06008408,2011YQ09000504,2012YQ12004407)国家高技术研究发展计划(2012AA020202) (2012AA020202)国家自然科学基金重点项目(20735005,21275159,31100591). (20735005,21275159,31100591)

A method of concatamers of Q peptides (QconCATs) protein labeled with ¹⁸O-multiple reaction monitoring mass spectrometry for absolute quantification of proteins is established. The purity of the QconCAT recombinant protein was characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and its purity was above 99%. The relative molecular mass was approximately 63.4 kDa. The peptides digested from the QconCAT recombinant protein and the extract of Thermoanaerobacter tengcongensis (TTE) were analyzed by mass spectrometry. The raw data were processed by pFind and pLabel softwares. The results showed that the efficiencies of protein digestion and the ¹⁸O labeling efficiency were able to meet the need of the protein quantification. The performance of the method was evaluated. The absolute contents of the selected proteins in TTE were determined with the relative standard deviations of less than 20% and the accuracy is high. The method not only avoid using the expensive reagent of stable isotope labeling with amino acids in cell culture (SILAC), but also provides an alternative way for the accurately absolute quantification of proteins in biological samples for quantitative proteomic research.
- concatamers of Q peptides (QconCATs),
- isotope dilution,
- multiple reaction monitoring mass spectrometry,
- ¹⁸O labeling,
- proteomics
1. [1]
  [1] Cao D, Zhang Y J, Qian X H. Journal of Chinese Mass Spectrometry Society (曹冬, 张养军, 钱小红. 质谱学报), 2008, 29(3): 185
2. [2]
  [2] Pratt J M, Simpson D M, Doherty M K, et al. Nat Protoc, 2006, 1(2): 1029
3. [3]
  [3] Beynon R J, Doherty M K, Pratt J M, et al. Nat Methods, 2005, 2(8): 587
4. [4]
  [4] Ding C, Li Y H, Kim B J, et al. J Proteome Res, 2011, 10(8): 3652
5. [5]
  [5] Swainston N, Jameson D, Carroll K. Proteomics, 2011, 11(2), 329
6. [6]
  [6] Rivers J, Simpson D M, Robertson D H L, et al. Mol Cell Proteomics, 2007, 6(8): 1416
7. [7]
  [7] Lin H J, Zhang A H, Cai Y, et al. Chemistry of Life (林虹君, 张爱红, 蔡耘, 等. 生命的化学), 2009, 29(6): 923
8. [8]
  [8] Liu W, He F C, Jiang Y. Chemistry of Life (刘伟, 贺福初, 姜颖. 生命的化学), 2009, 29(3): 427
9. [9]
  [9] Schiess R, Wollscheid B, Aebersold R. Mol Oncol, 2009, 3(1): 33
10. [10]
  [10] Picotti P, Aebersold R. Nat Methods, 2012, 9(6): 555
11. [11]
  [11] Gallien S, Duriez E, Domon B. J Mass Spectrom, 2011, 46(3): 298
12. [12]
  [12] Kiyonami R, Schoen A, Prakash A, et al. Mol Cell Proteomics, 2011, 10(2): 1
13. [13]
  [13] Jenkins R E, Kitteringham N R, Hunter C L, et al. Proteomics, 2006, 6(6): 1934
14. [14]
  [14] Zhao Y, Ying W T, Qian X H. Chemistry of Life (赵焱, 应万涛, 钱小红. 生命的化学), 2008, 28(2): 210
15. [15]
  [15] Zhao Y, Lu Z, Jia W, et al. Chinese Journal of Analytical Chemistry (赵焱, 卢庄, 贾伟, 等. 分析化学), 2010, 38(1): 91
16. [16]
  [16] Zhang J, Zhou L Q, Tian F, et al. Chinese Journal of Chromatography (张姣, 周廉淇, 田芳, 等. 色谱), 2013, 31(2): 102
17. [17]
  "
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]
  Yaling Chen . Basic Theory and Competitive Exam Analysis of Dynamic Isotope Effect. University Chemistry, 2024, 39(8): 403-410. doi: 10.3866/PKU.DXHX202311093
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]
  Ruolin CHENG , Haoran WANG , Jing REN , Yingying MA , Huagen LIANG . Efficient photocatalytic CO₂ cycloaddition over W₁₈O₄₉/NH₂-UiO-66 composite catalyst. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 523-532. doi: 10.11862/CJIC.20230349
5. [5]
  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
6. [6]
  Lingbang Qiu , Jiangmin Jiang , Libo Wang , Lang Bai , Fei Zhou , Gaoyu Zhou , Quanchao Zhuang , Yanhua Cui . 原位电化学阻抗谱监测长寿命热电池Nb₁₂WO₃₃正极材料的高温双放电机制. Acta Physico-Chimica Sinica, 2025, 41(5): 100040-. doi: 10.1016/j.actphy.2024.100040
7. [7]
  Zian Lin , Yingxue Jin . Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) for Disease Marker Screening and Identification: A Comprehensive Experiment Teaching Reform in Instrumental Analysis. University Chemistry, 2024, 39(11): 327-334. doi: 10.12461/PKU.DXHX202403066
8. [8]
  Tianlong Zhang , Rongling Zhang , Hongsheng Tang , Yan Li , Hua Li . Online Monitoring and Mechanistic Analysis of 3,5-diamino-1,2,4-triazole (DAT) Synthesis via Raman Spectroscopy: A Recommendation for a Comprehensive Instrumental Analysis Experiment. University Chemistry, 2024, 39(6): 303-311. doi: 10.3866/PKU.DXHX202312006
9. [9]
  Jianjun LI , Mingjie REN , Lili ZHANG , Lingling ZENG , Huiling WANG , Xiangwu MENG . UV-assisted degradation of tetracycline hydrochloride by MnFe₂O₄@activated carbon activated persulfate. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1869-1880. doi: 10.11862/CJIC.20240187
10. [10]
  Jinwang Wu , Qijing Xie , Chengliang Zhang , Haifeng Shi . 自旋极化增强ZnFe_1.2Co_0.8O₄/BiVO₄ S型异质结光催化性能降解四环素. Acta Physico-Chimica Sinica, 2025, 41(5): 100050-. doi: 10.1016/j.actphy.2025.100050
11. [11]
  Renxiao Liang , Zhe Zhong , Zhangling Jin , Lijuan Shi , Yixia Jia . A Palladium/Chiral Phosphoric Acid Relay Catalysis for the One-Pot Three-Step Synthesis of Chiral Tetrahydroquinoline. University Chemistry, 2024, 39(5): 209-217. doi: 10.3866/PKU.DXHX202311024
12. [12]
  Yixuan Zhu , Qingtong Wang , Jin Li , Lin Chen , Junlong Zhao . Blog of Oxytocin. University Chemistry, 2024, 39(9): 134-140. doi: 10.12461/PKU.DXHX202310090
13. [13]
  Endong YANG , Haoze TIAN , Ke ZHANG , Yongbing LOU . Efficient oxygen evolution reaction of CuCo₂O₄/NiFe-layered bimetallic hydroxide core-shell nanoflower sphere arrays. Chinese Journal of Inorganic Chemistry, 2024, 40(5): 930-940. doi: 10.11862/CJIC.20230369
14. [14]
  Danqing Wu , Jiajun Liu , Tianyu Li , Dazhen Xu , Zhiwei Miao . Research Progress on the Simultaneous Construction of C—O and C—X Bonds via 1,2-Difunctionalization of Olefins through Radical Pathways. University Chemistry, 2024, 39(11): 146-157. doi: 10.12461/PKU.DXHX202403087
15. [15]
  Hao Wu , Zhen Liu , Dachang Bai . ¹H NMR Spectrum of Amide Compounds. University Chemistry, 2024, 39(3): 231-238. doi: 10.3866/PKU.DXHX202309020
16. [16]
  Yukun Xing , Xiaoyu Xie , Fangfang Chen . A Sunlit Gift: Vitamin D. University Chemistry, 2024, 39(9): 28-34. doi: 10.12461/PKU.DXHX202402006
17. [17]
  Donghui PAN , Yuping XU , Xinyu WANG , Lizhen WANG , Junjie YAN , Dongjian SHI , Min YANG , Mingqing CHEN . Preparation and in vivo tracing of ⁶⁸Ga-labeled PM_2.5 mimetic particles for positron emission tomography imaging. Chinese Journal of Inorganic Chemistry, 2024, 40(4): 669-676. doi: 10.11862/CJIC.20230468
18. [18]
  Lina Guo , Ruizhe Li , Chuang Sun , Xiaoli Luo , Yiqiu Shi , Hong Yuan , Shuxin Ouyang , Tierui Zhang . 层状双金属氢氧化物的层间阴离子对衍生的Ni-Al₂O₃催化剂光热催化CO₂甲烷化反应的影响. Acta Physico-Chimica Sinica, 2025, 41(1): 2309002-. doi: 10.3866/PKU.WHXB202309002
19. [19]
  Jiandong Liu , Zhijia Zhang , Mikhail Kamenskii , Filipp Volkov , Svetlana Eliseeva , Jianmin Ma . Research Progress on Cathode Electrolyte Interphase in High-Voltage Lithium Batteries. Acta Physico-Chimica Sinica, 2025, 41(2): 100011-. doi: 10.3866/PKU.WHXB202308048
20. [20]
  Jin Tong , Shuyan Yu . Crystal Engineering for Supramolecular Chirality. University Chemistry, 2024, 39(3): 86-93. doi: 10.3866/PKU.DXHX202308113

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(276)
HTML views(46)

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

A novel method for absolute protein quantification using 18O isotope labeled concatamers of Q peptides combined with isotope dilution-multiple reaction monitoring mass spectrometry

Metrics

通讯作者: 陈斌, bchen63@163.com

A novel method for absolute protein quantification using ¹⁸O isotope labeled concatamers of Q peptides combined with isotope dilution-multiple reaction monitoring mass spectrometry