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

TY - JOUR

T1 - Analysis of phosphoinositide 3-kinase inhibitors by bottom-up electron-transfer dissociation hydrogen/deuterium exchange mass spectrometry

AU - Masson, Glenn R.

AU - Maslen, Sarah L.

AU - Williams, Roger L.

N1 - Funding Information: The work was supported by AstraZeneca/LMB Blue Sky Initiative [MC-A024-5PF9G to R.L.W.] and the Medical Research Council [MC-U105184308 to R.L.W.]. G.R.M. was supported by a Henslow Research Fellowship from The Cambridge Philosophical Society and St Catharine's College, Cambridge. Publisher Copyright: © 2017 The Author(s).

PY - 2017/5/16

Y1 - 2017/5/16

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85019910003

U2 - 10.1042/BCJ20170127

DO - 10.1042/BCJ20170127

M3 - Article

C2 - 28381646

AN - SCOPUS:85019910003

SN - 0264-6021

VL - 474

SP - 1867

EP - 1877

JO - Biochemical Journal

JF - Biochemical Journal

IS - 11

ER -