Identification of gel-separated tumor marker proteins by mass spectrometry.
Two-dimensional gel electrophoresis with subsequent analysis by mass spectrometry was applied to study differences in protein expression between benign and malignant solid tumors from human beast, lung and ovary cells. Cells from freshly resected clinical material were lysed and the extracts were subjected to isoelectric focusing with immobilized pH gradients followed by second-dimensional separation on 10-13% sodium dodecyl sulfate (SDS)/polyacrylamide gels. Polypeptides were identified using matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry after in-gel protein digestion. Some of the upregulated polypeptides in malignant cells are of potential importance as markers of tumor proliferation. Twenty such proteins were identified, ten constituting novel identifications and ten sequence verifications of previously gel-matched proteins. The proteins identified span a wide range of functions, but several cases of protein truncation were found. Truncated forms of cytokeratins 6D and 8, and of cathepsin D were identified. Truncated froms of these over-expressed proteins support the presence of proteolytic processing steps in tumor material. The protein processing and the difference between protein and mRNA abundancies in tumors of different malignancy and origin suggest that studies at the protein level are important for an understanding of tumor phenotypes.
Bergman
A C
AC
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Benjamin
T
T
Alaiya
A
A
Waltham
M
M
Sakaguchi
K
K
Franzén
B
B
Linder
S
S
Bergman
T
T
Auer
G
G
Appella
E
E
Wirth
P J
PJ
Jörnvall
H
H
D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000737:chemistry; D002457:Cell Extracts / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D010051:Ovarian Neoplasms / Q000737:chemistry
D014408:Biomarkers, Tumor; D002457:Cell Extracts