title: The intermediate filament protein, vimentin, in the lens is a target for cross-linking by transglutaminase.
abstract: Mere addition of Ca2+ to a lens cortical homogenate (bovine) generates a series of products composed of a variety of high molecular weight vimentin species. The Ca2+-induced cross-linking of this cytoskeletal element seems to be mediated by the intrinsic transglutaminase of lens, because the reaction could be blocked at the monomeric state of vimentin by the inclusion of small synthetic substrates of the enzyme dansylcadaverine or dansyl-epsilon-aminocaproyl-Gln-Gln-Ile-Val. These compounds are known to compete against the Gln or Lys functionalities of proteins that would participate in forming the Nepsilon(gamma-glutamyl)lysine protein-to-protein cross-links. The cytosolic transglutaminase-catalyzed reactions could be reproduced with purified bovine lens vimentin and also with recombinant human vimentin preparations. Employing the latter system, we have titrated the transglutaminase-reactive sites of vimentin and, by sequencing the dansyl-tracer-labeled segments of the protein, we have shown that residues Gln453 and Gln460 served as acceptor functionalities and Lys97, Lys104, Lys294, and Lys439 as electron donor functionalities in vimentin. The transglutaminase-dependent reaction of this intermediate filament protein might influence the shape and plasticity of the fiber cells, and the enzyme-catalyzed cross-linking of vimentin, in conjunction with other lens constituents, may contribute to the process of cataract formation.
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
authors.forename: S
authors.initials: S
authors.lastname: Clément
authors.forename: P T
authors.initials: PT
authors.lastname: Velasco
authors.forename: S N
authors.initials: SN
authors.lastname: Murthy
authors.forename: J H
authors.initials: JH
authors.lastname: Wilson
authors.forename: T J
authors.initials: TJ
authors.lastname: Lukas
authors.forename: R D
authors.initials: RD
authors.lastname: Goldman
authors.forename: L
authors.initials: L
authors.lastname: Lorand
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002103:Cadaverine / Q000031:analogs & derivatives / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003599:Cytoskeleton / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D011503:Transglutaminases / Q000378:metabolism*; D014746:Vimentin / Q000378:metabolism*
chemical_list: D014746:Vimentin; D005973:Glutamine; D011503:Transglutaminases; C008542:monodansylcadaverine; D008239:Lysine; D002103:Cadaverine; D002118:Calcium