biostudies-literatureScietti LDH | NIHR | Health Services and Delivery Research Programme (Health Services and Delivery Research (HS&DR) Programme)Fondazione CariploEuropean Commission (EC)Giovanni Armenise-Harvard FoundationDH | NIHR | Health Services and Delivery Research Programme (Health Services and Delivery ResearchMinistero dell'Istruzione, dell'Università e della RicercaNational Institute for Health Research (NIHR)Fondazione Italiana per la Ricerca sul CancroFondazione Cariplo (Cariplo Foundation)European CommissionFondazione Italiana per la Ricerca sul Cancro (Italian Foundation for Cancer Research)Ministero dell'Istruzione, dell'Università e della Ricerca (Ministry of Education, University and Research)3163https://www.ebi.ac.uk/biostudies/studies/S-EPMC6082870biostudies-literatureUnknown9(1)Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. Malfunctions in these enzymes cause severe connective tissue disorders. Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. Our understanding of LH3 functions is currently hampered by lack of molecular structure information. Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. Known disease-related mutations map in close proximity to the catalytic sites. Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases.Nature communicationsMolecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3.PMC6082870MSCA-IF Grant agreement n. 745934 -“COME TRUE” (id. 2015-0768)My First AIRC Grant (Grant id. 20075)MSCA-IF Grant agreement n. 745934 –FP7/2007-201 (Biostruct-X) Grant agreements 7551 and 10205Programma Giovani Ricercatori Rita Levi-MontalciniCDA 2013Programma Dipartimenti di Eccellenza 2018-202214/21/45"COME TRUE" (id. 2015-0768)Banushi BFumagalli MGiulotto EDe Giorgi FNergadze SKhoriauli LForneris FScietti LCucca LProfumo AOlieric VChiapparino ABasu SGissen PfalseMolecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3.Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. Malfunctions in these enzymes cause severe connective tissue disorders. Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. Our understanding of LH3 functions is currently hampered by lack of molecular structure information. Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. Known disease-related mutations map in close proximity to the catalytic sites. Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases.2018-01-01T00:00:00Z2018 Aug2024-11-20T09:41:01.213Z2019-03-26T23:56:31ZS-EPMC60828703008981210.1038/s41467-018-05631-5