Mechanical strain rapidly redistributes tyrosine phosphorylated proteins in human intestinal Caco-2 cells.
Repetitive strain stimulates proliferation and modulates differentiation in human Caco-2 intestinal epithelial cells via tyrosine kinase activity. We therefore sought to characterize strain modulation of tyrosine phosphorylation in Caco-2 cells. Immunoblotting for phosphotyrosine demonstrated that repetitive strain (10 cpm, 10% strain) rapidly increased tyrosine phosphorylation of 125-, 70-, 60-, and 50-kDa bands in the soluble fraction by 94+/-31, 145+/-21, 365+/-46, and 1240+/-240%, respectively (p<0.05, n=4). However, strain decreased tyrosine phosphorylated band intensity of the 125-, 70-, 60-, and 50-kDa proteins in the particulate fraction by 81+/-17, 70+/-23, 79+/-7, and 59+/-23%, respectively (p<0.05, n=4). The decreased band intensity in the particulate fraction was not due to decreased tyrosine kinase activity because strain equally increased tyrosine kinase activity in both soluble and particulate fractions. Cyclic strain at a physiologically relevant amplitude and frequency appears to modulate the subcellular distribution of tyrosine phosphorylated proteins in human Caco-2 intestinal cells.
Han
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O
Departments of Surgery, Yale University School of Medicine and the Connecticut VA Health Care System, New Haven, Connecticut, 06520-8062, USA.
Sumpio
B E
BE
Basson
M D
MD
D018938:Caco-2 Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007422:Intestines / Q000201:enzymology / Q000473:pathology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D013314:Stress, Mechanical; D014443:Tyrosine / Q000378:metabolism
D011506:Proteins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases