title: N-Glycosylation pattern of the zymogenic form of human matrix metalloproteinase-9.
abstract: Glycosylation of proteins has profound consequences on the activities of macromolecules and their interactions with inhibitors/substrates. Matrix metalloproteinase-9 (MMP-9, also known as gelatinase B) is a member of the MMP family of zinc-dependent endopeptidases, with critical functions in both physiological and pathological processes. MMP-9, a glycosylated MMP, is implicated in inflammation, angiogenesis and tumor metastasis. We have determined by the use of mass spectrometry that of the three possible N-glycosylation sites in human MMP-9 only two are glycosylated. The N-glycosylation sites are at asparagines in positions 38 and 120, the first site within the propeptide domain of the zymogenic form (pro-MMP-9) of the enzyme and the second in the catalytic domain. The chemical nature of the sugar attachments to both these sites was determined by mass spectrometry. Both N-glycosylation sites have NeuAcalpha(1,2)-Galbeta(1,4)-GlcNAcbeta(1,2)-Manalpha(1,3)-[NeuAcalpha(1,2)-Galbeta(1,4)-GlcNAcbeta(1,2)-Manalpha(1,6)-]Manbeta(1,4)-GlcNAcbeta(1,4)-[Fucalpha(1,6)-]GlcNAcbeta oligosaccharide chains. A computational model of glycosylated pro-MMP-9 was generated and it was studied by dynamics simulations
authors.affiliation: Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.
authors.forename: Lakshmi P
authors.initials: LP
authors.lastname: Kotra
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Rafael
authors.initials: R
authors.lastname: Fridman
authors.forename: Ron
authors.initials: R
authors.lastname: Orlando
authors.forename: Shahriar
authors.initials: S
authors.lastname: Mobashery
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D020134:Catalytic Domain; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002918:Chymotrypsin / Q000378:metabolism; D003198:Computer Simulation; D004792:Enzyme Precursors / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
chemical_list: D004792:Enzyme Precursors; D007527:Isoenzymes; D006026:Glycoside Hydrolases; D002918:Chymotrypsin; D014357:Trypsin; D020780:Matrix Metalloproteinase 9