PMID- 19401150
OWN - NLM
STAT- MEDLINE
DCOM- 20090610
LR  - 20131121
IS  - 1878-5875 (Electronic)
IS  - 1357-2725 (Linking)
VI  - 41
IP  - 7
DP  - 2009 Jul
TI  - Differences in hydroxylation and binding of Notch and HIF-1alpha demonstrate 
      substrate selectivity for factor inhibiting HIF-1 (FIH-1).
PG  - 1563-71
LID - 10.1016/j.biocel.2009.01.005 [doi]
AB  - FIH-1, factor inhibiting hypoxia-inducible factor-1 (HIF-1), regulates oxygen 
      sensing by hydroxylating an asparagine within HIF-alpha. It also hydroxylates 
      asparagines in many proteins containing ankyrin repeats, including Notch1-3, p105 
      and I?B?. Relative binding affinity and hydroxylation rate are crucial 
      determinants of substrate selection and modification. We determined the 
      contributions of substrate sequence composition and length and of oxygen 
      concentration to the FIH-1-binding and/or hydroxylation of Notch1-4 and compared 
      them with those for HIF-1alpha. We also demonstrated hydroxylation of two 
      asparagines in Notch2 and 3, corresponding to Sites 1 and 2 of Notch1, by mass 
      spectrometry for the first time. Our data demonstrate that substrate length has a 
      much greater influence on FIH-1-dependent hydroxylation of Notch than of 
      HIF-1alpha, predominantly through binding affinity rather than maximal reaction 
      velocity. The K(m) value of FIH-1 for Notch1, < 0.2 microM, is at least 250-fold 
      lower than that of 50 microM for HIF-1alpha. Site 1 of Notch1-3 appeared the 
      preferred site of FIH-1 hydroxylation in these substrates. Interestingly, binding 
      of Notch4 to FIH-1 was observed with an affinity almost 10-fold lower than for 
      Notch1-3, but no hydroxylation was detected. Importantly, we demonstrate that the 
      K(m) of FIH-1 for oxygen at the preferred Site 1 of Notch1-3, 10-19 microM, is an 
      order of magnitude lower than that for Site 2 or HIF-1alpha. Hence, at least 
      during in vitro hydroxylation, Notch is likely to become efficiently hydroxylated 
      by FIH-1 even under relatively severe hypoxic conditions, where HIF-1alpha 
      hydroxylation would be reduced.
FAU - Wilkins, Sarah E
AU  - Wilkins SE
AD  - School of Molecular and Biomedical Science and the Australian Research Council 
      Special Research Centre for the Molecular Genetics of Development, University of 
      Adelaide, Adelaide, SA 5005, Australia.
FAU - Hyvarinen, Jaana
AU  - Hyvarinen J
FAU - Chicher, Johana
AU  - Chicher J
FAU - Gorman, Jeffrey J
AU  - Gorman JJ
FAU - Peet, Daniel J
AU  - Peet DJ
FAU - Bilton, Rebecca L
AU  - Bilton RL
FAU - Koivunen, Peppi
AU  - Koivunen P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090120
PL  - Netherlands
TA  - Int J Biochem Cell Biol
JT  - The international journal of biochemistry & cell biology
JID - 9508482
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Peptides)
RN  - 0 (Receptors, Notch)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Repressor Proteins)
RN  - 7006-34-0 (Asparagine)
RN  - EC 1.- (Mixed Function Oxygenases)
RN  - EC 1.14.11.- (HIF1AN protein, human)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Asparagine/metabolism
MH  - Humans
MH  - Hydroxylation
MH  - Hypoxia-Inducible Factor 1, alpha Subunit/*metabolism
MH  - Kinetics
MH  - Mice
MH  - Mixed Function Oxygenases
MH  - Molecular Sequence Data
MH  - Oxygen/metabolism
MH  - Peptides/chemistry/metabolism
MH  - Protein Binding
MH  - Receptors, Notch/chemistry/*metabolism
MH  - Recombinant Proteins/metabolism
MH  - Repressor Proteins/*metabolism
MH  - Substrate Specificity
EDAT- 2009/04/30 09:00
MHDA- 2009/06/11 09:00
CRDT- 2009/04/30 09:00
PHST- 2008/11/06 00:00 [received]
PHST- 2009/01/08 00:00 [revised]
PHST- 2009/01/12 00:00 [accepted]
PHST- 2009/04/30 09:00 [entrez]
PHST- 2009/04/30 09:00 [pubmed]
PHST- 2009/06/11 09:00 [medline]
AID - S1357-2725(09)00009-0 [pii]
AID - 10.1016/j.biocel.2009.01.005 [doi]
PST - ppublish
SO  - Int J Biochem Cell Biol. 2009 Jul;41(7):1563-71. doi: 
      10.1016/j.biocel.2009.01.005. Epub 2009 Jan 20.