Profiling human brain proteome by multi-dimensional separations coupled with MS
Young Mok Park
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorJin Young Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorKyung-Hoon Kwon
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorSang Kwang Lee
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorYoung Hye Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorSe-Young Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorGun Wook Park
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorJeong Hwa Lee
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorBonghee Lee
Department of Anatomy and Neurobiology, College of Medicine, Cheju National University, Jeju, Republic of Korea
Search for more papers by this authorCorresponding Author
Jong Shin Yoo Dr.
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Mass Spectrometer Development Team, Korea Basic Science Institute, 52 Yeoeun-dong, Yusung-gu, Daejeon 305–333, Republic of Korea Fax: +82-42-865-3419===Search for more papers by this authorYoung Mok Park
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorJin Young Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorKyung-Hoon Kwon
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorSang Kwang Lee
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorYoung Hye Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorSe-Young Kim
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorGun Wook Park
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorJeong Hwa Lee
Proteomics Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Search for more papers by this authorBonghee Lee
Department of Anatomy and Neurobiology, College of Medicine, Cheju National University, Jeju, Republic of Korea
Search for more papers by this authorCorresponding Author
Jong Shin Yoo Dr.
Mass Spectrometer Development Team, Korea Basic Science Institute, Daejeon, Republic of Korea
Mass Spectrometer Development Team, Korea Basic Science Institute, 52 Yeoeun-dong, Yusung-gu, Daejeon 305–333, Republic of Korea Fax: +82-42-865-3419===Search for more papers by this authorAbstract
In our initial attempt to analyze the human brain proteome, we applied multi-dimensional protein separation and identification techniques using a combination of sample fractionation, 1-D SDS-PAGE, and MS analysis. The complexity of human brain proteome requires multiple fractionation strategies to extend the range and total number of proteins identified. According to the method of Klose (Methods Mol. Biol. 1999, 112, 67), proteins of the temporal lobe of human brain were fractionated into (i) cytoplasmic and nucleoplasmic, (ii) membrane and other structural, and (iii) DNA-binding proteins. Each fraction was then separated by SDS-PAGE, and the resulting gel line was cut into approximately 50 bands. After trypsin digestion, the resulting peptides from each band were analyzed by RP-LC/ESI-MS/MS using an LTQ spectrometer. The SEQUEST search program, which searched against the IPI database, was used for peptide sequence identification, and peptide sequences were validated by reversed sequence database search and filtered by the Protein Hit Score. Ultimately, 1533 proteins could be detected from the human brain. We classified the identified proteins according to their distribution on cellular components. Among these proteins, 24% were membrane proteins. Our results show that the multiple separation strategy is effective for high-throughput characterization of proteins from complex proteomic mixtures.
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Citing Literature
Special Issue:THE HUPO BRAIN PROTEOME PROJECT Concerted Proteome Analysis of the Brain
No. 18 September 2006
Pages 4978-4986
