title: TIMP-3 binds to sulfated glycosaminoglycans of the extracellular matrix.
abstract: Of the four known tissue inhibitors of metalloproteinases (TIMPs), TIMP-3 is distinguished by its tighter binding to the extracellular matrix. The present results show that glycosaminoglycans such as heparin, heparan sulfate, chondroitin sulfates A, B, and C, and sulfated compounds such as suramin and pentosan efficiently extract TIMP-3 from the postpartum rat uterus. Enzymatic treatment by heparinase III or chondroitinase ABC also releases TIMP-3, but neither one alone gives complete release. Confocal microscopy shows colocalization of heparan sulfate and TIMP-3 in the endometrium subjacent to the lumen of the uterus. Immunostaining of TIMP-3 is lost upon digestion of tissue sections with heparinase III and chondroitinase ABC. The N-terminal domain of human TIMP-3 was expressed and found to bind to heparin with affinity similar to that of full-length mouse TIMP-3. The A and B beta-strands of the N-terminal domain of TIMP-3 contain two potential heparin-binding sequences rich in lysine and arginine; these strands should form a double track on the outer surface of TIMP-3. Synthetic peptides corresponding to segments of these two strands compete for heparin in the DNase II binding assay. TIMP-3 binding may be important for the cellular regulation of activity of the matrix metalloproteinases.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Miami, Florida 33101, USA.
authors.forename: W H
authors.initials: WH
authors.lastname: Yu
authors.forename: S
authors.initials: S
authors.lastname: Yu
authors.forename: Q
authors.initials: Q
authors.lastname: Meng
authors.forename: K
authors.initials: K
authors.lastname: Brew
authors.forename: J F
authors.initials: JF
authors.lastname: Woessner
mesh_terms: D000478:Alkylation; D000818:Animals; D019765:Chondroitin ABC Lyase / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D003871:Dermatan Sulfate / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004717:Endometrium / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D015723:Gene Library; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D018613:Microscopy, Confocal; D010455:Peptides / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000378:metabolism; D049590:Postpartum Period; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*; D013498:Suramin / Q000378:metabolism; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014599:Uterus / Q000378:metabolism
chemical_list: D019765:Chondroitin ABC Lyase; D002809:Chondroitin Sulfates; D018076:DNA, Complementary; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D010455:Peptides; D011133:Polysaccharide-Lyases; D011994:Recombinant Proteins; D013455:Sulfur; D013498:Suramin; D019717:Tissue Inhibitor of Metalloproteinase-3; C021349:heparitinsulfate lyase