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.
Stubbs
Christopher J
CJ
Department of Chemistry and Oxford Centre for Integrative Systems Biology, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
Loenarz
Christoph
C
Mecinović
Jasmin
J
Yeoh
Kar Kheng
KK
Hindley
Nicola
N
Liénard
Benoît M
BM
Sobott
Frank
F
Schofield
Christopher J
CJ
Flashman
Emily
E
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