Analysis of phosphoinositide 3-kinase inhibitors by bottom-up electron-transfer dissociation hydrogen/deuterium exchange mass spectrometry

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 th...

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Bibliographic Details
Published in:	Biochemical journal 2017-06, Vol.474 (11), p.1867-1877
Main Authors:	Masson, Glenn R, Maslen, Sarah L, Williams, Roger L
Format:	Article
Language:	eng
Subjects:	Antineoplastic Agents - chemistry Antineoplastic Agents - metabolism Antineoplastic Agents - pharmacology Binding Sites Class I Phosphatidylinositol 3-Kinases Class Ia Phosphatidylinositol 3-Kinase - chemistry Class Ia Phosphatidylinositol 3-Kinase - genetics Class Ia Phosphatidylinositol 3-Kinase - metabolism Deuterium Exchange Measurement Drug Evaluation, Preclinical - methods Electron Transport Enzyme Inhibitors - chemistry Enzyme Inhibitors - metabolism Enzyme Inhibitors - pharmacology Humans Indazoles - chemistry Indazoles - metabolism Indazoles - pharmacology Models, Molecular Molecular Weight Oligonucleotides - antagonists & inhibitors Oligonucleotides - chemistry Oligonucleotides - genetics Oligonucleotides - metabolism Peptide Fragments - antagonists & inhibitors Peptide Fragments - chemistry Peptide Fragments - genetics Peptide Fragments - metabolism Phosphatidylinositol 3-Kinases - chemistry Phosphatidylinositol 3-Kinases - genetics Phosphatidylinositol 3-Kinases - metabolism Phosphoinositide-3 Kinase Inhibitors Protein Conformation Purines - chemistry Purines - metabolism Purines - pharmacology Pyridazines Quinazolinones - chemistry Quinazolinones - metabolism Quinazolinones - pharmacology Quinolines - chemistry Quinolines - metabolism Quinolines - pharmacology Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - metabolism Reproducibility of Results Signal Processing, Computer-Assisted Sulfonamides - chemistry Sulfonamides - metabolism Sulfonamides - pharmacology Tandem Mass Spectrometry Triazines - chemistry Triazines - metabolism Triazines - pharmacology
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Summary:	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.
ISSN:	0264-6021 1470-8728