title: Ambidextrous binding of cell and membrane bilayers by soluble matrix metalloproteinase-12.
abstract: Matrix metalloproteinases (MMPs) regulate tissue remodelling, inflammation and disease progression. Some soluble MMPs are inexplicably active near cell surfaces. Here we demonstrate the binding of MMP-12 directly to bilayers and cellular membranes using paramagnetic NMR and fluorescence. Opposing sides of the catalytic domain engage spin-labelled membrane mimics. Loops project from the β-sheet interface to contact the phospholipid bilayer with basic and hydrophobic residues. The distal membrane interface comprises loops on the other side of the catalytic cleft. Both interfaces mediate MMP-12 association with vesicles and cell membranes. MMP-12 binds plasma membranes and is internalized to hydrophobic perinuclear features, the nuclear membrane and inside the nucleus within minutes. While binding of TIMP-2 to MMP-12 hinders membrane interactions beside the active site, TIMP-2-inhibited MMP-12 binds vesicles and cells, suggesting compensatory rotation of its membrane approaches. MMP-12 association with diverse cell membranes may target its activities to modulate innate immune responses and inflammation.
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, Missouri 65211, USA.
authors.forename: Rama K
authors.initials: RK
authors.lastname: Koppisetti
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, Missouri 65211, USA.
authors.forename: Yan G
authors.initials: YG
authors.lastname: Fulcher
authors.affiliation: Molecular Cytology Core, 120 Bond Life Sciences Center, University of Missouri, Columbia, Missouri 65211, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Jurkevich
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, Missouri 65211, USA.
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Prior
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, Missouri 65211, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Xu
authors.affiliation: College of Medical and Dental Sciences, School of Cancer Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors.forename: Marc
authors.initials: M
authors.lastname: Lenoir
authors.affiliation: College of Medical and Dental Sciences, School of Cancer Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Overduin
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, Missouri 65211, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Van Doren
mesh_terms: D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D006367:HeLa Cells; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology; D053508:Matrix Metalloproteinase 12 / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D013113:Spin Labels; D055672:Static Electricity; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism*
chemical_list: D019812:Heparan Sulfate Proteoglycans; D008051:Lipid Bilayers; D046912:Multiprotein Complexes; D010743:Phospholipids; D013113:Spin Labels; C560823:TIMP2 protein, human; D019716:Tissue Inhibitor of Metalloproteinase-2; D053508:Matrix Metalloproteinase 12