Differential expression of phosphatidylethanol-amine-binding protein in rat hepatoma cell lines: analyses of tumor necrosis factor-alpha-resistant cKDH-8/11 and -sensitive KDH-8/YK cells by two-dimensional gel electrophoresis.
To determine intracellular factors influencing the sensitivity of cancer cells to tumor necrosis factor-alpha (TNF-alpha), we studied the expression of intracellular proteins in TNF-alpha-resistant cKDH-8/11 and -sensitive KDH-8/YK rat hepatoma cell lines using the technique of two-dimensional gel electrophoresis (2-DE). From the 2-DE patterns, it was demonstrated that TNF-alpha-resistant cKDH-8/11 cells had increased levels of protein of molecular weight (Mr) 22 500 and isoelectric point (pI) 5.2, compared with TNF-alpha-sensitive KDH-8/YK cells. Therefore, we excised cyanogen bromide (CNBr) fragments of proteins in the spot for N-terminal sequencing. Microsequencing for the CNBr fragments identified the protein as rat phosphatidylethanolamine-binding protein. These findings suggest that the intracellular phosphatidylethanolamine-binding protein could be one of the factors responsible for the resistance of cKDH-8/11 cells to TNF-alpha-induced cell death.
Kuramitsu
Y
Y
Department of Biochemistry, Yamaguchi University School of Medicine, Ube, Japan.
Fujimoto
M
M
Tanaka
T
T
Ohata
J
J
Nakamura
K
K
D000595:Amino Acid Sequence; D000727:Androgen-Binding Protein; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008969:Molecular Sequence Data; D049294:Phospholipid Transfer Proteins; D060789:Prostatein; D051381:Rats; D060765:Secretoglobins; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014598:Uteroglobin; D060788:Mammaglobin B
D000727:Androgen-Binding Protein; D002352:Carrier Proteins; D049294:Phospholipid Transfer Proteins; D060789:Prostatein; D060765:Secretoglobins; D014409:Tumor Necrosis Factor-alpha; D014598:Uteroglobin; C511484:Scgb1d2 protein, rat; C511483:Scgb1d4 protein, rat; C556468:Scgb2a1 protein, rat; D060788:Mammaglobin B