title: Quantitative proteomics analysis of the cAMP/protein kinase A signaling pathway.
abstract: To define the proteins whose expression is regulated by cAMP and protein kinase A (PKA), we used a quantitative proteomics approach in studies of wild-type (WT) and kin- (PKA-null) S49 murine T lymphoma cells. We also compared the impact of endogenous increases in the level of cAMP [by forskolin (Fsk) and the phosphodiesterase inhibitor isobutylmethylxanthine (IBMX)] or by a cAMP analogue (8-CPT-cAMP). We identified 1056 proteins in WT and kin- S49 cells and found that 8-CPT-cAMP and Fsk with IBMX produced differences in protein expression. WT S49 cells had a correlation coefficient of 0.41 between DNA microarray data and the proteomics analysis in cells incubated with 8-CPT-cAMP for 24 h and a correlation coefficient of 0.42 between the DNA microarray data obtained at 6 h and the changes in protein expression after incubation with 8-CPT-cAMP for 24 h. Glutathione reductase (Gsr) had a higher level of basal expression in kin- S49 cells than in WT cells. Consistent with this finding, kin- cells are less sensitive to cell killing and generation of malondialdehyde than are WT cells incubated with H(2)O(2). Cyclic AMP acting via PKA thus has a broad impact on protein expression in mammalian cells, including in the regulation of Gsr and oxidative stress.
authors.affiliation: Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, California 92093-0654, United States.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Guo
authors.forename: Andrea
authors.initials: A
authors.lastname: Wilderman
authors.forename: Lingzhi
authors.initials: L
authors.lastname: Zhang
authors.forename: Susan S
authors.initials: SS
authors.lastname: Taylor
authors.forename: Paul A
authors.initials: PA
authors.lastname: Insel
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D008315:Malondialdehyde / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D040901:Proteomics; D015398:Signal Transduction
chemical_list: D008315:Malondialdehyde; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases