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Patel BB , Li XM , Dixon MP , Blagoi EL , Seeholzer SH , Chen Y , Miller CG , He YA , Tetruashvily M , Chaudhry AH , Ke E , Xie J , Cooper H , Bellacosa A , Clapper ML , Boman BM , Zhang T , Litwin S , Ross EA , Conrad P , Crowell JA , Kopelovich L , Knudson A , Yeung AT
Searchable high-resolution 2D gel proteome of the human colon crypt
J Proteome Res. 2007 Jun;6(6) :2232-8
PMID: 17444668   
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Abstract
We seek alterations in protein patterns at the earliest possible step on the path to cancer, namely, in cells of the target tissue from normal persons versus the corresponding normally appearing cells from persons who are heterozygous for mutation in a tumor suppressor gene that predisposes strongly to carcinoma in that tissue. To begin a systematic comparison of the proteomes of cells from normal and from neoplastic colons, we have undertaken the isolation of human colon crypts that are derived from the normal-appearing mucosa of left (descending) colon of patients with sporadic colorectal cancer. Two-dimensional (2D) gel electrophoresis is a proteomic approach that excels in the resolution of protein isoforms. Here, we document the practicality of this approach with human samples using gels of three overlapping pH ranges. For the first time, about 800 nonredundant proteins and 900 isoforms from purified human colonic crypts were identified, permitting an assessment of the contributions of protein isoforms. These interactive, searchable, hyperlink-enabled proteome maps and gene ontology analyses will facilitate future studies to discover the earliest markers and intervention targets during progression to colon cancer.
Notes
Patel, Bhavinkumar B Li, Xin-Ming Dixon, Maketa P Blagoi, Elena L Seeholzer, Steven H Chen, Yibai Miller, C Glenn He, Yin A Tetruashvily, Mazell Chaudhry, Anam H Ke, Eileen Xie, Joan Cooper, Harry Bellacosa, Alfonso Clapper, Margie L Boman, Bruce M Zhang, Tao Litwin, Samuel Ross, Eric A Conrad, Peggy Crowell, James A Kopelovich, Levy Knudson, Alfred Yeung, Anthony T CA119242/CA/NCI NIH HHS/United States DK063014/DK/NIDDK NIH HHS/United States N01-NC-15103/PHS HHS/United States P30CA06927/CA/NCI NIH HHS/United States Comparative Study Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States Journal of proteome research J Proteome Res. 2007 Jun;6(6):2232-8. Epub 2007 Apr 20.