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Image Search Results
Journal: The Journal of Biological Chemistry
Article Title: The yeast AMP-activated protein kinase Snf1 phosphorylates the inositol polyphosphate kinase Kcs1
doi: 10.1016/j.jbc.2024.105657
Figure Lengend Snippet: Kcs1 is a phosphoprotein that undergoes Snf1-dependent phosphorylation in vivo . A , diagram of the inositol polyphosphate biosynthetic pathway. The boat conformation for InsP 3 is shown below a flat representation of InsP 3 with the one and six positions of inositol labeled. Proteins catalyzing each reaction are indicated. B , mass spectrometry identifies decreased phosphorylation of Kcs1 Ser 646 under steady-state conditions of nitrogen limitation in a strain with a kinase-defective allele of snf1 . For quantitative mass spectrometry by SILAC, both strains for analysis contained deletions of lys1 and arg4 and were grown in media with either normal Lys and Arg or in media with the indicated stable isotopic forms. The wild-type strain is deleted for snf1 and carries a low-copy centromeric plasmid containing the wild-type SNF1 sequence with native promoter. The plasmid in the test strain encodes a kinase-defective allele of snf1 with the indicated substitution of Arg for Lys 84. The posterior error probability (PEP) is the probability that the observed peptide-spectrum match for identification of the protein is incorrect. The ratio of phosphorylated peptide in the mutant and wild-type strains, normalized against total protein, is indicated. C , mass spectrometry of a wild-type strain of the filamentous Σ1278b background grown under conditions of nitrogen limitation identified phosphorylation of Kcs1 at Ser residues 537 and 646. The probability of correct amino acid identification is indicated, along with confidence and false discovery rate.
Article Snippet: For Stable Isotope Labeling by
Techniques: Phospho-proteomics, In Vivo, Labeling, Mass Spectrometry, Multiplex sample analysis, Plasmid Preparation, Sequencing, Mutagenesis