title: The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases. Role of the fibronectin type II-like repeats.
abstract: Several matrix metalloproteinases, including the 92-kDa and 72-kDa gelatinases, macrophage metalloelastase (MME), and matrilysin degrade insoluble elastin. Because elastolytically active MME and matrilysin consist only of a catalytic domain (CD), we speculated that the homologous CDs of the 92-kDa and 72-kDa gelatinases would confer their elastolytic activities. In contrast to the MME CD, the 92 and 72 CDs expressed in Escherichia coli (lacking the internal fibronectin type II-like repeats) had no elastase activity, although both were gelatinolytic and cleaved a thiopeptolide substrate at rates comparable to the full-length gelatinases. To test the role of the fibronectin type II-like repeats in elastolytic activity, we expressed the 92-kDa gelatinase CD with its fibronectin type II-like repeats (92 CD/FN) in yeast. 92 CD/FN degraded insoluble elastin with activity comparable to full-length 92-kDa gelatinase. 92 and 72 CDs lacking the fibronectin type II-like repeats did not bind elastin, whereas the parent enzymes and 92 CD/FN did bind elastin. Furthermore, recombinant 92-kDa fibronectin type II-like repeats inhibited binding of the 92-kDa gelatinase to elastin. We conclude that the 92- and 72-kDa gelatinases require the fibronectin type II-like repeats for elastase activity.
authors.affiliation: Respiratory and Critical Care Division, Washington University School of Medicine at Jewish Hospital, St. Louis, Missouri 63110, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Shipley
authors.forename: G A
authors.initials: GA
authors.lastname: Doyle
authors.forename: C J
authors.initials: CJ
authors.lastname: Fliszar
authors.forename: Q Z
authors.initials: QZ
authors.lastname: Ye
authors.forename: L L
authors.initials: LL
authors.lastname: Johnson
authors.forename: S D
authors.initials: SD
authors.lastname: Shapiro
authors.forename: H G
authors.initials: HG
authors.lastname: Welgus
authors.forename: R M
authors.initials: RM
authors.lastname: Senior
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers; D004549:Elastin; D004926:Escherichia coli; D005353:Fibronectins / Q000737:chemistry*; D018093:Gelatinases / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008264:Macrophages / Q000201:enzymology; D020783:Matrix Metalloproteinase 7; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000737:chemistry* / Q000378:metabolism*; D010843:Pichia; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; D005353:Fibronectins; D011994:Recombinant Proteins; D004549:Elastin; D010196:Pancreatic Elastase; D018093:Gelatinases; D008666:Metalloendopeptidases; D020783:Matrix Metalloproteinase 7