title: Protein disulfide isomerase-mediated disulfide bonds regulate the gelatinolytic activity and secretion of matrix metalloproteinase-9.
abstract: Matrix metalloproteinase-9 (MMP-9) is one of the major MMPs that can degrade extracellular matrix. Besides normal physiological functions, MMP-9 is involved in metastasis and tumor angiogenesis. Although several inhibitors of MMP-9 have been identified, in vivo regulators of MMP-9 activation are unknown. In the present study we intended to investigate novel therapeutic target protein(s) that regulate MMP-9 activation and/or secretion. We have identified protein disulfide isomerase as a novel upstream regulator of MMP-9. Mass spectrometric analysis of post-translational modification in MMP-9 confirmed six disulfide bonds in the catalytic domain and one disulfide bond in the hemopexin domain of MMP-9. Establishment of cells that overexpressed wild-type and mutant forms of MMP-9 revealed that 'cysteine-switch' and disulfide bonds within the catalytic domain are necessary for the secretion and intracellular trafficking of MMP-9. However, the disulfide bond of the hemopexin domain and other cysteines have no significant role in secretion. These insights into the secretion of MMP-9 constitute the basis for the development of potential drugs against metastasis.
authors.affiliation: Chemical Library Validation Team, Chemical Biology Core Facility, Chemical Biology Department, RIKEN Advanced Science Institute, Saitama, Japan.
authors.forename: Maola M G
authors.initials: MM
authors.lastname: Khan
authors.forename: Siro
authors.initials: S
authors.lastname: Simizu
authors.forename: Takehiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Akiko
authors.initials: A
authors.lastname: Masuda
authors.forename: Makoto
authors.initials: M
authors.lastname: Kawatani
authors.forename: Makoto
authors.initials: M
authors.lastname: Muroi
authors.forename: Naoshi
authors.initials: N
authors.lastname: Dohmae
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Osada
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D002460:Cell Line; D003545:Cysteine; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000235:genetics / Q000473:pathology; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D034622:RNA Interference; D034741:RNA, Small Interfering
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D021382:Protein Sorting Signals; D034741:RNA, Small Interfering; D020780:Matrix Metalloproteinase 9; D019704:Protein Disulfide-Isomerases; D003545:Cysteine