Collagen lysyl hydroxylases (LH1-3) are Fe2+- and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations that promote fibrosis and cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 Å crystal structure and X-ray scattering on dimer assemblies for the LH domain of L230 in Acanthamoeba polyphaga mimivirus. Loop residues in the double-stranded β-helix core generate a tail-to-tail dimer. A stabilizing hydrophobic leucine locks into an aromatic tyrosine-pocket on the opposite subunit. An active site triad coordinates Fe2+. The two active sites flank a deep surface cleft that suggest dimerization creates a collagen-binding site. Loss of Fe2+-binding disrupts the dimer. Dimer disruption and charge reversal in the cleft increase K m and reduce LH activity. Ectopic L230 expression in tumors promotes collagen cross-linking and metastasis. These insights suggest inhibitor targets for fibrosis and cancer.
|State||Published - Dec 1 2018|
Bibliographical noteFunding Information:
This work was supported in part by the National Institutes of Health grants R21AR060978 (M.Y.), R01CA105155 (J.M.K. and M.Y.), P50CA70907 (J.M.K.); Cancer Prevention and Research Institute of Texas (CPRIT) grant RP160652 (J.M.K.); The International Association for the Study of Lung Cancer Postdoctoral Fellowship (H.-F.G.); the Elza A. and Ina S. Freeman Professorship in Lung Cancer (J.M.K.); and The Welch Foundation F-1390 (K.N.D.); NIH GM123252 (K.N.D.); and CPRIT RP160657 (K.N.D.). We thank Kathryn Brunett and the SIBYLS beamline staff for collecting SAXS data. The SIBYLS beamline is supported through the Integrated Diffraction Analysis Technologies Program, which is supported by Department of Energy Office of Biological and Environmental Research, National Institute of Health project MINOS (R01GM105404), and a High-End Instrumentation Grant S10OD018483. We thank Andre Mueller from Wyatt Technology for MALS data analysis support.
© 2018 The Author(s).
ASJC Scopus subject areas
- Chemistry (all)
- Biochemistry, Genetics and Molecular Biology (all)
- Physics and Astronomy (all)