Proteome analysis of focal adhesion kinase (FAK)-overexpressing cells.
We established several focal adhesion kinase (FAK) cDNA-transfected cells and found that FAK-transfected HL-60 (HL-60/FAK) cells are resistant to apoptosis induced with hydrogen peroxide, etoposide and radiation compared with the parental HL-60 or the vector-transfected (HL-60/Vect) cells. We carried out proteome analysis to study the mechanism of resistance to apoptosis in HL-60/FAK cells. Among 300 spots resolved in two-dimensional gels, ca. 10% of them were significantly increased in HL-60/FAK cells compared with HL-60/Vect cells, whereas ca. 2% of them were decreased or disappeared. These proteins were performed for further analysis by Western blots or N-terminal sequencing or mass spectrometry. Increased proteins included stress proteins such as hsp90, ribosomal proteins, and antioxidant enzymes such as peroxyredoxin 2. Some of these proteins are assumed to contribute to the antiapoptotic action of FAK.
Utsubo
Ryoko
R
Department of Biochemistry, Kyoritsu University of Pharmacy, Tokyo, Japan.
Sonoda
Yoshiko
Y
Takahashi
Reiko
R
Iijima
Shiro
S
Aizu-Yokota
Eriko
E
Kasahara
Tadashi
T
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D020869:Gene Expression Profiling; D018922:HL-60 Cells; D006801:Humans; D013058:Mass Spectrometry; D016133:Polymerase Chain Reaction; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis*; D020543:Proteome / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013879:Thioredoxins / Q000096:biosynthesis; D014162:Transfection
D020543:Proteome; D012333:RNA, Messenger; D013879:Thioredoxins; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human