PMID- 16952279
OWN - NLM
STAT- MEDLINE
DCOM- 20070129
LR  - 20220330
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 401
IP  - 1
DP  - 2007 Jan 1
TI  - Studies on the activity of the hypoxia-inducible-factor hydroxylases using an 
      oxygen consumption assay.
PG  - 227-34
AB  - The activity and levels of the metazoan HIF (hypoxia-inducible factor) are 
      regulated by its hydroxylation, catalysed by 2OG (2-oxoglutarate)- and 
      Fe(II)-dependent dioxygenases. An oxygen consumption assay was developed and used 
      to study the relationship between HIF hydroxylase activity and oxygen 
      concentration for recombinant forms of two human HIF hydroxylases, PHD2 (prolyl 
      hydroxylase domain-containing protein 2) and FIH (factor inhibiting HIF), and 
      compared with two other 2OG-dependent dioxygenases. Although there are caveats on 
      the absolute values, the apparent K(m) (oxygen) values for PHD2 and FIH were 
      within the range observed for other 2OG oxygenases. Recombinant protein 
      substrates were found to have lower apparent K(m) (oxygen) values compared with 
      shorter synthetic peptides of HIF. The analyses also suggest that human PHD2 is 
      selective for fragments of the C-terminal over the N-terminal oxygen-dependent 
      degradation domain of HIF-1alpha. The present results, albeit obtained under 
      non-physiological conditions, imply that the apparent K(m) (oxygen) values of the 
      HIF hydroxylases enable them to act as oxygen sensors providing their in vivo 
      capacity is appropriately matched to a hydroxylation-sensitive signalling 
      pathway.
FAU - Ehrismann, Dominic
AU  - Ehrismann D
AD  - Chemistry Research Laboratory, Department of Chemistry and Oxford Centre for 
      Molecular Sciences, University of Oxford, Oxford OXI 3TA, UK.
FAU - Flashman, Emily
AU  - Flashman E
FAU - Genn, David N
AU  - Genn DN
FAU - Mathioudakis, Nicolas
AU  - Mathioudakis N
FAU - Hewitson, Kirsty S
AU  - Hewitson KS
FAU - Ratcliffe, Peter J
AU  - Ratcliffe PJ
FAU - Schofield, Christopher J
AU  - Schofield CJ
LA  - eng
GR  - BBS/B/07683/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
GR  - WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (Transcription Factors)
RN  - EC 1.- (Mixed Function Oxygenases)
RN  - EC 1.1.3.4 (Glucose Oxidase)
RN  - EC 1.14.11.- (HIF1AN protein, human)
RN  - EC 1.14.11.2 (EGLN1 protein, human)
RN  - EC 1.14.11.2 (Procollagen-Proline Dioxygenase)
RN  - EC 1.14.11.29 (Hypoxia-Inducible Factor-Proline Dioxygenases)
SB  - IM
MH  - Cloning, Molecular
MH  - Glucose Oxidase/metabolism
MH  - Hypoxia-Inducible Factor-Proline Dioxygenases
MH  - Kinetics
MH  - Mixed Function Oxygenases
MH  - *Oxygen Consumption
MH  - Peptide Fragments/chemistry/metabolism
MH  - Procollagen-Proline Dioxygenase/genetics/*metabolism
MH  - Protein Binding
MH  - Recombinant Proteins/metabolism
MH  - Repressor Proteins/genetics/metabolism
MH  - Substrate Specificity
MH  - Transcription Factors/genetics/metabolism
PMC - PMC1698668
EDAT- 2006/09/06 09:00
MHDA- 2007/01/30 09:00
PMCR- 2007/07/01
CRDT- 2006/09/06 09:00
PHST- 2006/09/06 09:00 [pubmed]
PHST- 2007/01/30 09:00 [medline]
PHST- 2006/09/06 09:00 [entrez]
PHST- 2007/07/01 00:00 [pmc-release]
AID - BJ20061151 [pii]
AID - bj4010227 [pii]
AID - 10.1042/BJ20061151 [doi]
PST - ppublish
SO  - Biochem J. 2007 Jan 1;401(1):227-34. doi: 10.1042/BJ20061151.