title: Bioavailable affinity label for collagen prolyl 4-hydroxylase.
abstract: Collagen is the most abundant protein in animals. Its prevalent 4-hydroxyproline residues contribute greatly to its conformational stability. The hydroxyl groups arise from a post-translational modification catalyzed by the nonheme iron-dependent enzyme, collagen prolyl 4-hydroxylase (P4H). Here, we report that 4-oxo-5,6-epoxyhexanoate, a mimic of the α-ketoglutarate co-substrate, inactivates human P4H. The inactivation installs a ketone functionality in P4H, providing a handle for proteomic experiments. Caenorhabditis elegans exposed to the esterified epoxy ketone displays the phenotype of a worm lacking P4H. Thus, this affinity label can be used to mediate collagen stability in an animal, as is desirable in the treatment of a variety of fibrotic diseases.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706-1544, USA.
authors.forename: James D
authors.initials: JD
authors.lastname: Vasta
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Higgin
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Kersteen
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Raines
mesh_terms: D000818:Animals; D055162:Biocatalysis; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007659:Ketones / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D064828:Prolyl Hydroxylases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D007659:Ketones; D003094:Collagen; D064828:Prolyl Hydroxylases