The structure of the second CysD domain of MUC2 and role in mucin organization by transglutaminase-based cross-linking

Christian V Recktenwald; Göran Karlsson; Maria-Jose Garcia-Bonete; Gergely Katona; Maja Jensen; Richard Lymer; Malin Bäckström; Malin E V Johansson; Gunnar C Hansson; Sergio Trillo-Muyo

doi:10.1016/j.celrep.2024.114207

The structure of the second CysD domain of MUC2 and role in mucin organization by transglutaminase-based cross-linking

Cell Rep. 2024 May 9;43(5):114207. doi: 10.1016/j.celrep.2024.114207. Online ahead of print.

Affiliations

¹ Department of Medical Biochemistry and Cell Biology, University of Gothenburg, 405 30 Gothenburg, Sweden. Electronic address: christian.recktenwald@medkem.gu.se.
² Swedish NMR Centre, University of Gothenburg, 405 30 Gothenburg, Sweden.
³ Department of Medical Biochemistry and Cell Biology, University of Gothenburg, 405 30 Gothenburg, Sweden.
⁴ Department of Chemistry and Molecular Biology, University of Gothenburg, 405 30 Gothenburg, Sweden.
⁵ Mammalian Protein Expression core facility, University of Gothenburg, 405 30 Gothenburg, Sweden.
⁶ Department of Medical Biochemistry and Cell Biology, University of Gothenburg, 405 30 Gothenburg, Sweden. Electronic address: gunnar.hansson@medkem.gu.se.
⁷ Department of Medical Biochemistry and Cell Biology, University of Gothenburg, 405 30 Gothenburg, Sweden. Electronic address: sergio.muyo@medkem.gu.se.

PMID: 38733585
DOI: 10.1016/j.celrep.2024.114207

Abstract

The MUC2 mucin protects the colonic epithelium by a two-layered mucus with an inner attached bacteria-free layer and an outer layer harboring commensal bacteria. CysD domains are 100 amino-acid-long sequences containing 10 cysteines that separate highly O-glycosylated proline, threonine, serine (PTS) regions in mucins. The structure of the second CysD, CysD2, of MUC2 is now solved by nuclear magnetic resonance. CysD2 shows a stable stalk region predicted to be partly covered by adjacent O-glycans attached to neighboring PTS sequences, whereas the CysD2 tip with three flexible loops is suggested to be well exposed. It shows transient dimer interactions at acidic pH, weakened at physiological pH. This transient interaction can be stabilized in vitro and in vivo by transglutaminase 3-catalyzed isopeptide bonds, preferring a specific glutamine residue on one flexible loop. This covalent dimer is modeled suggesting that CysD domains act as connecting hubs for covalent stabilization of mucins to form a protective mucus.

Keywords: 3D structure; CP: Microbiology; CP: Molecular biology; TGM3; bacteria; colitis; colon; intestine; isopeptide bond; mucin; mucus; transglutaminase.