title: MMP-12 catalytic domain recognizes and cleaves at multiple sites in human skin collagen type I and type III.
abstract: Collagens of either soft connective or mineralized tissues are subject to continuous remodeling and turnover. Undesired cleavage can be the result of an imbalance between proteases and their inhibitors. Owing to their superhelical structure, collagens are resistant to many proteases and matrix metalloproteinases (MMPs) are required to initiate further degradation by other enzymes. Several MMPs are known to degrade collagens, but the action of MMP-12 has not yet been studied in detail. In this work, the potential of MMP-12 in recognizing sites in human skin collagen types I and III has been investigated. The catalytic domain of MMP-12 binds to the triple helix and cleaves the typical sites -Gly(775)-Leu(776)- in alpha-2 type I collagen and -Gly(775)-Ile(776)- in alpha-1 type I and type III collagens and at multiple other sites in both collagen types. Moreover, it was observed that the region around these typical sites contains comparatively less prolines, of which some have been proven to be only partially hydroxylated. This is of relevance since partial hydroxylation in the vicinity of a potential scissile bond may have a local effect on the conformational thermodynamics with probable consequences on the collagenolysis process. Taken together, the results of the present work confirm that the catalytic domain of MMP-12 alone binds and degrades collagens I and III.
authors.affiliation: Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
authors.forename: Samuel
authors.initials: S
authors.lastname: Taddese
authors.forename: Michael C
authors.initials: MC
authors.lastname: Jung
authors.forename: Christian
authors.initials: C
authors.lastname: Ihling
authors.forename: Andrea
authors.initials: A
authors.lastname: Heinz
authors.forename: Reinhard H H
authors.initials: RH
authors.lastname: Neubert
authors.forename: Christian E H
authors.initials: CE
authors.lastname: Schmelzer
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D020134:Catalytic Domain; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D024042:Collagen Type I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000090602:Collagen Type I, alpha 1 Chain; D024061:Collagen Type III / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D066298:In Vitro Techniques; D053508:Matrix Metalloproteinase 12 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D013816:Thermodynamics
chemical_list: C439971:COL3A1 protein, human; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D024061:Collagen Type III; D010446:Peptide Fragments; D011994:Recombinant Proteins; D003094:Collagen; C507122:MMP12 protein, human; D053508:Matrix Metalloproteinase 12