Analyzing alkaline proteins in human colon crypt proteome

Journal of Proteome Research
Xin-Ming LiAnthony T Yeung

Abstract

Normal human colon crypt protein extract was resolved by two-dimensional gel electrophoresis using pH 6-11 immobilized pH gradient strips in the first dimension. The optimized isoelectric focusing protocol includes cup-loading sample application at the anode and 1.2% hydroxyethyl disulfide (DeStreak), 15% 2-propanol and 5% glycerol in the rehydration buffer. Spots were well resolved across the entire pH range up to 11. A total of 311 protein spots were identified by mass spectrometry and peptide mass mapping. After combining isoforms, 231 nonredundant proteins were grouped into 16 categories according to their subcellular locations, and 17 categories according to their physiological functions. Histone proteins, ribosomal proteins and mitochondrial proteins were among the well-resolved highest p/ proteins. Application of this protocol to the analysis of normal and neoplastic colon crypts will contribute to the proteomic study of colorectal tumorigenesis since a significant portion of the human proteins is in basic pH range.

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Citations

Jan 19, 2010·Molecular & Cellular Proteomics : MCP·Svetlana E NikoulinaSteven W Taylor
Jan 13, 2009·Pancreas·Eileen KeAnthony T Yeung
Dec 25, 2013·Laboratory Investigation; a Journal of Technical Methods and Pathology·Michael K DameD Kim Turgeon
Jan 22, 2011·Science China. Life Sciences·Wei MiXiaoHong Qian
Feb 24, 2006·Proteomics·Thibault ChazeBrigitte Picard
Oct 17, 2009·Proteomics·Lena KaskBengt Bjellqvist
Mar 24, 2006·Molecular & Cellular Proteomics : MCP·Xuezhi BiChoy-Leong Hew
Dec 12, 2013·Pancreas·Kathryn T ChenAnthony T Yeung
Jan 29, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·S M Shahjahan MiahKerry S Campbell
Jul 4, 2018·International Journal of Molecular Sciences·Andrea AntonosanteAnnamaria Cimini
Mar 18, 2011·Oncotarget·Bhavinkumar B PatelLevy Kopelovich
Apr 30, 2017·Clinical and Experimental Pharmacology & Physiology·Yi-Ping Huang, Nai-Wen Chang

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