Protein pyrophosphorylation by diphosphoinositol pentakisphosphate (InsP7).
Diphosphoinositol polyphosphates, also known as inositol pyrophosphates, are a family of water soluble inositol phosphates that possess diphosphate or pyrophosphate moieties. In the presence of divalent cations such as Mg(2+), the "high energy" beta phosphate can be transferred from the inositol pyrophosphates, InsP(7) and InsP(8), to prephosphorylated serine residues on proteins, to form pyrophosphoserine. This chapter provides detailed methods to identify proteins that are substrates for pyrophosphorylation by InsP(7), conduct phosphorylation assays on purified protein, and detect protein pyrophosphorylation.
Werner
J Kent
JK
The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Speed
Traci
T
Bhandari
Rashna
R
D000595:Amino Acid Sequence; D001345:Autoradiography / Q000379:methods; D001671:Biochemistry / Q000379:methods*; D015153:Blotting, Western / Q000379:methods; D011756:Diphosphates / Q000032:analysis* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007295:Inositol Phosphates / Q000032:analysis* / Q000378:metabolism; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D011756:Diphosphates; D007295:Inositol Phosphates; D011506:Proteins; D011993:Recombinant Fusion Proteins; C402123:inositol heptakisphosphate; C079849:1-diphosphoinositol pentakisphosphate