8ONE

Crystal Structure of full-length Human Lysyl Hydroxylase LH3 - Asp190Ser mutant - Cocrystal with Fe2+, Mn2+, UDP-Glucose

Summary for 8ONE

• Entry DOI: 10.2210/pdb8one/pdb
• Descriptor: Multifunctional procollagen lysine hydroxylase and glycosyltransferase LH3, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, 2-OXOGLUTARIC ACID, ... (9 entities in total)
• Functional Keywords: collagen biosynthesis; extracellular matrix; post-translational modifications; collagen glycosylations; glycosyltransferase; lysyl hydroxylase, transferase
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 1
• Total formula weight: 84614.37

Authors

Mattoteia, D.,De Marco, M.,Pinnola, A.,Faravelli, S.,Scietti, L.,Forneris, F. (deposition date: 2023-04-02, release date: 2023-07-19, Last modification date: 2024-11-13)

Primary citation

Mattoteia, D.,Chiapparino, A.,Fumagalli, M.,De Marco, M.,De Giorgi, F.,Negro, L.,Pinnola, A.,Faravelli, S.,Roscioli, T.,Scietti, L.,Forneris, F.
Identification of Regulatory Molecular "Hot Spots" for LH/PLOD Collagen Glycosyltransferase Activity.
Int J Mol Sci, 24:-, 2023

PubMed Abstract: Hydroxylysine glycosylations are post-translational modifications (PTMs) essential for the maturation and homeostasis of fibrillar and non-fibrillar collagen molecules. The multifunctional collagen lysyl hydroxylase 3 (LH3/PLOD3) and the collagen galactosyltransferase GLT25D1 are the human enzymes that have been identified as being responsible for the glycosylation of collagen lysines, although a precise description of the contribution of each enzyme to these essential PTMs has not yet been provided in the literature. LH3/PLOD3 is thought to be capable of performing two chemically distinct collagen glycosyltransferase reactions using the same catalytic site: an inverting beta-1,O-galactosylation of hydroxylysines (Gal-T) and a retaining alpha-1,2-glucosylation of galactosyl hydroxylysines (Glc-T). In this work, we have combined indirect luminescence-based assays with direct mass spectrometry-based assays and molecular structure studies to demonstrate that LH3/PLOD3 only has Glc-T activity and that GLT25D1 only has Gal-T activity. Structure-guided mutagenesis confirmed that the Glc-T activity is defined by key residues in the first-shell environment of the glycosyltransferase catalytic site as well as by long-range contributions from residues within the same glycosyltransferase (GT) domain. By solving the molecular structures and characterizing the interactions and solving the molecular structures of human LH3/PLOD3 in complex with different UDP-sugar analogs, we show how these studies could provide insights for LH3/PLOD3 glycosyltransferase inhibitor development. Collectively, our data provide new tools for the direct investigation of collagen hydroxylysine PTMs and a comprehensive overview of the complex network of shapes, charges, and interactions that enable LH3/PLOD3 glycosyltransferase activities, expanding the molecular framework and facilitating an improved understanding and manipulation of glycosyltransferase functions in biomedical applications.

PubMed: 37446392
DOI: 10.3390/ijms241311213

Experimental method

X-RAY DIFFRACTION (2.3 Å)