Searchable high-resolution 2D gel proteome of the human colon crypt.
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.
Patel
Bhavinkumar B
BB
Division of Basic Science, Medical Science, and Population Science, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
Li
Xin-Ming
XM
Dixon
Maketa P
MP
Blagoi
Elena L
EL
Seeholzer
Steven H
SH
Chen
Yibai
Y
Miller
C Glenn
CG
He
Yin A
YA
Tetruashvily
Mazell
M
Chaudhry
Anam H
AH
Ke
Eileen
E
Xie
Joan
J
Cooper
Harry
H
Bellacosa
Alfonso
A
Clapper
Margie L
ML
Boman
Bruce M
BM
Zhang
Tao
T
Litwin
Samuel
S
Ross
Eric A
EA
Conrad
Peggy
P
Crowell
James A
JA
Kopelovich
Levy
L
Knudson
Alfred
A
Yeung
Anthony T
AT
D002374:Catalase / Q000032:analysis; D003106:Colon / Q000737:chemistry*; D015179:Colorectal Neoplasms / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000592:standards; D005982:Glutathione Transferase / Q000032:analysis; D006801:Humans; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D013482:Superoxide Dismutase / Q000032:analysis
D011506:Proteins; D020543:Proteome; D002374:Catalase; D013482:Superoxide Dismutase; D005982:Glutathione Transferase