1. Mannose binding lectin is hydroxylated by collagen prolyl-4-hydroxylase and inhibited by some PHD inhibitors
- Author
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Maxwell, Patrick, Bhute, Vijesh, Harte, James, Houghton, Jack, Maxwell, Patrick [0000-0002-0338-2679], and Apollo - University of Cambridge Repository
- Subjects
hypoxia inducible factor ,mannose binding lectin ,post-translational ,collagen prolyl-4-hydroxylase ,Procollagen-Proline Dioxygenase ,protein processing ,chemical and pharmacologic phenomena ,Prolyl-Hydroxylase Inhibitors ,bacterial infections and mycoses ,Prolyl Hydroxylases ,PHD inhibitors ,Hypoxia-Inducible Factor-Proline Dioxygenases ,HEK293 cells ,Collagen ,FG-4592 ,chronic kidney disease ,renal anemia ,vadadustat ,molidustat - Abstract
Background Mannose binding lectin (MBL) is an important component of innate immune defence. MBL undergoes oligomerization to generate high molecular weight (HMW) forms which act as pattern recognition molecules to detect and opsonize various microorganisms. Several post-translational modifications including prolyl hydroxylation are known to affect the oligomerization of MBL. Yet, the enzyme(s) which hydroxylate proline in the collagen-like domain residues have not been identified and the significance of prolyl hydroxylation is incompletely understood. Methods To investigate post-translational modifications of MBL, we stably expressed Myc-DDK tagged MBL in HEK293S cells. We used pharmacological and genetic inhibition of 2-oxoglutarate dependent dioxygenases (2OGDD) to identify the enzyme required for prolyl hydroxylation of MBL. We performed mass spectrometry to determine the effects of various inhibitors on MBL modifications. Results Secretion of HMW MBL was impaired by inhibitors of the superfamily of 2OGDD, and was dependent on prolyl-4-hydroxylase subunit alpha 1. Roxadustat and vadadustat, but not molidustat, led to significant suppression of hydroxylation and secretion of HMW forms of MBL. Conclusions These data suggest that prolyl hydroxylation in the collagen-like domain of MBL is mediated by collagen prolyl 4 hydroxylase. Reduced MBL activity is likely to be an off-target effect of some, but not all PHD inhibitors. There may be advantages in selective PHD inhibitors that would not interfere with MBL production.
- Published
- 2020