title: Analysis of phosphoinositide 3-kinase inhibitors by bottom-up electron-transfer dissociation hydrogen/deuterium exchange mass spectrometry.
abstract: Until recently, one of the major limitations of hydrogen/deuterium exchange mass spectrometry (HDX-MS) was the peptide-level resolution afforded by proteolytic digestion. This limitation can be selectively overcome through the use of electron-transfer dissociation to fragment peptides in a manner that allows the retention of the deuterium signal to produce hydrogen/deuterium exchange tandem mass spectrometry (HDX-MS/MS). Here, we describe the application of HDX-MS/MS to structurally screen inhibitors of the oncogene phosphoinositide 3-kinase catalytic p110α subunit. HDX-MS/MS analysis is able to discern a conserved mechanism of inhibition common to a range of inhibitors. Owing to the relatively minor amounts of protein required, this technique may be utilised in pharmaceutical development for screening potential therapeutics.
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Cambridge CB2 0QH, U.K.
authors.forename: Glenn R
authors.initials: GR
authors.lastname: Masson
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Cambridge CB2 0QH, U.K.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Maslen
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Cambridge CB2 0QH, U.K. gmasson@mrc-lmb.cam.ac.uk.
authors.forename: Roger L
authors.initials: RL
authors.lastname: Williams
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001665:Binding Sites; D058534:Class I Phosphatidylinositol 3-Kinases; D058543:Class Ia Phosphatidylinositol 3-Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D041961:Deuterium Exchange Measurement; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004579:Electron Transport; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007191:Indazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D008958:Models, Molecular; D008970:Molecular Weight; D009841:Oligonucleotides / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011487:Protein Conformation; D011687:Purines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011724:Pyridazines; D052999:Quinazolinones / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011804:Quinolines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted; D013449:Sulfonamides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D053719:Tandem Mass Spectrometry; D014227:Triazines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000096926:Benzenesulfonamides
chemical_list: D000970:Antineoplastic Agents; D058534:Class I Phosphatidylinositol 3-Kinases; D058543:Class Ia Phosphatidylinositol 3-Kinase; D004791:Enzyme Inhibitors; D007191:Indazoles; D009841:Oligonucleotides; D010446:Peptide Fragments; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011687:Purines; D011724:Pyridazines; D052999:Quinazolinones; D011804:Quinolines; D011993:Recombinant Fusion Proteins; D013449:Sulfonamides; D014227:Triazines; C532162:2-(1H-indazol-4-yl)-6-(4-methanesulfonylpiperazin-1-ylmethyl)-4-morpholin-4-ylthieno(3,2-d)pyrimidine; C510150:ZSTK474; C561454:omipalisib; D000096926:Benzenesulfonamides; C588001:PIK3R1 protein, human; C484760:PIK3CA protein, human; C552946:idelalisib
keywords: ETD; HDX-MS; HDX-MS/MS; PI3K