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Biochemical and Structural Insights into FIH-Catalysed Hydroxylation of Transient Receptor Potential Ankyrin Repeat Domains |
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| Authors: | Benjamin G Saward Thomas M Leissing Ian J Clifton Anthony Tumber Christopher M Timperley Richard J Hopkinson Prof Christopher J Schofield |
| Affiliation: | 1. Department of Chemistry and the, Ineos Oxford Institute for Antimicrobial Research, Chemistry Research Laboratory, Mansfield Road, University of Oxford, Oxford, OX1 3TA UK
These authors contributed equally to this work.;2. Department of Chemistry and the, Ineos Oxford Institute for Antimicrobial Research, Chemistry Research Laboratory, Mansfield Road, University of Oxford, Oxford, OX1 3TA UK;3. CBR Division, Defence Science and Technology Laboratory (DSTL), Porton Down, Salisbury, SP4 0JQ UK |
| Abstract: | Transient receptor potential (TRP) channels have important roles in environmental sensing in animals. Human TRP subfamily A member 1 (TRPA1) is responsible for sensing allyl isothiocyanate (AITC) and other electrophilic sensory irritants. TRP subfamily vanilloid member 3 (TRPV3) is involved in skin maintenance. TRPV3 is a reported substrate of the 2-oxoglutarate oxygenase factor inhibiting hypoxia-inducible factor (FIH). We report biochemical and structural studies concerning asparaginyl hydroxylation of the ankyrin repeat domains (ARDs) of TRPA1 and TRPV3 catalysed by FIH. The results with ARD peptides support a previous report on FIH-catalysed TRPV3 hydroxylation and show that, of the 12 potential TRPA1 sequences investigated, one sequence (TRPA1 residues 322–348) undergoes hydroxylation at Asn336. Structural studies reveal that the TRPA1 and TRPV3 ARDs bind to FIH with a similar overall geometry to most other reported FIH substrates. However, the binding mode of TRPV3 to FIH is distinct from that of other substrates. |
| Keywords: | ankyrin factor inhibiting HIF (FIH) hypoxia inducible factor HIF oxygenases demethylases post translational modifications transient receptor potential (TRP) channel |