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Determination and comparison of specific activity of the HIF‐prolyl hydroxylases
Author(s) -
Tuckerman Jason R.,
Zhao Yuguang,
Hewitson Kirsty S.,
Tian Ya-Min,
Pugh Christopher W.,
Ratcliffe Peter J.,
Mole David R.
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.09.005
Subject(s) - hydroxylation , enzyme , hypoxia (environmental) , biochemistry , hypoxia inducible factors , recombinant dna , chemistry , substrate (aquarium) , microbiology and biotechnology , biology , oxygen , gene , ecology , organic chemistry
Hypoxia‐inducible factor (HIF) is a transcriptional complex that is regulated by oxygen sensitive hydroxylation of its α subunits by the prolyl hydroxylases PHD1, 2 and 3. To better understand the role of these enzymes in directing cellular responses to hypoxia, we derived an assay to determine their specific activity in both native cell extracts and recombinant sources of enzyme. We show that all three are capable of high rates of catalysis, in the order PHD2=PHD3 >PHD1, using substrate peptides derived from the C‐terminal degradation domain of HIF‐α subunits, and that each demonstrates similar and remarkable sensitivity to oxygen, commensurate with a common role in signaling hypoxia.