title: Enzymatic processing by MMP-2 and MMP-9 of wild-type and mutated mouse β-dystroglycan.
abstract: Dystroglycan (DG) is a membrane-associated protein complex formed by two noncovalently linked subunits, α-DG, a highly glycosylated extracellular protein, and β-DG, a transmembrane protein. The interface between the two DG subunits, which is crucial to maintain the integrity of the plasma membrane, involves the C-terminal domain of α-DG and the N-terminal extracellular domain of β-DG. It is well known that under both, physiological and pathological conditions, gelatinases (i.e. MMP-9 and/or MMP-2) can degrade DG, disrupting the connection between the extracellular matrix and the cytoskeleton. However, the molecular mechanisms and the exact cleavage sites underlying these events are still largely unknown. In a previous study, we have characterized the enzymatic digestion of the murine β-DG ectodomain by gelatinases, identifying a main cleavage site on the β-DG ectodomain produced by MMP-9. In this article, we have deepened the pattern of the β-DG ectodomain digestion by MMP-2 by using a combined approach based on SDS-PAGE, Orbitrap, and HPLC-ESI-IT mass spectrometry. Furthermore, we have characterized the kineticparameters of the digestion of some β-DG ectodomain mutants by gelatinases.
authors.affiliation: Dipartimento di Scienze Cliniche e Medicina Traslazionale, Università di Roma Tor Vergata, Roma, Italy.
authors.forename: Diego
authors.initials: D
authors.lastname: Sbardella
authors.forename: Rosanna
authors.initials: R
authors.lastname: Inzitari
authors.forename: Federica
authors.initials: F
authors.lastname: Iavarone
authors.forename: Magda
authors.initials: M
authors.lastname: Gioia
authors.forename: Stefano
authors.initials: S
authors.lastname: Marini
authors.forename: Francesca
authors.initials: F
authors.lastname: Sciandra
authors.forename: Massimo
authors.initials: M
authors.lastname: Castagnola
authors.forename: Philippe E
authors.initials: PE
authors.lastname: Van den Steen
authors.forename: Ghislain
authors.initials: G
authors.lastname: Opdenakker
authors.forename: Bruno
authors.initials: B
authors.lastname: Giardina
authors.forename: Andrea
authors.initials: A
authors.lastname: Brancaccio
authors.forename: Massimo
authors.initials: M
authors.lastname: Coletta
authors.forename: Manuela
authors.initials: M
authors.lastname: Bozzi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D049030:Dystroglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D059748:Proteolysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells
chemical_list: D010446:Peptide Fragments; D011994:Recombinant Proteins; D049030:Dystroglycans; D020778:Matrix Metalloproteinase 2; C522362:Mmp2 protein, mouse; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse