Application of a proteolysis/mass spectrometry method for investigating the effects of inhibitors on hydroxylase structure.
Limited proteolysis coupled to matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analyses can be used to screen for compounds that alter protein structure by monitoring stabilizing/destabilizing effects with respect to the rate and nature of proteolysis. When applied to prolyl hydroxylase 2, a key enzyme involved in human oxygen sensing, the method efficiently revealed differential effects on proteolytic stability for structurally similar compounds and for different substrates.
Department of Chemistry and Oxford Centre for Integrative Systems Biology, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
Christopher J
CJ
Stubbs
Christoph
C
Loenarz
Jasmin
J
Mecinović
Kar Kheng
KK
Yeoh
Nicola
N
Hindley
Benoît M
BM
Liénard
Frank
F
Sobott
Christopher J
CJ
Schofield
Emily
E
Flashman
D000818:Animals; D020134:Catalytic Domain / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000737:chemistry / Q000378:metabolism; D020128:Inhibitory Concentration 50; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D010446:Peptide Fragments / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004791:Enzyme Inhibitors; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010446:Peptide Fragments; D006899:Mixed Function Oxygenases