title: The C-terminal domain of tissue inhibitor of metalloproteinases-2 is required for cell binding but not for antimetalloproteinase activity.
abstract: We have generated a C-terminally-truncated form of recombinant tissue inhibitor of metalloproteinases-2 (designated rTIMP-2 delta) in which the region of the inhibitor extending from residue 128 to 194 and including 3 of the 6 disulfide bonds is deleted. rTIMP-2 and rTIMP-2 delta had similar inhibitory activities toward interstitial collagenase and inhibited the activation of the precursor form of matrix metalloproteinase-2 (proMMP-2). rTIMP-2 also bound with high affinity (Kd 0.99 nM) to HT1080 human fibrosarcoma cells treated with 12-O-tetradecanoyl-phorbol-13-acetate. However deletion of the C-terminal domain of TIMP-2 significantly lowered the cell surface binding affinity, with competition experiments indicating a 2 order of magnitude difference between rTIMP-2 and rTIMP-2 delta in the concentrations needed to displace 125I-labeled rTIMP-2 binding. These data indicate that the C-terminal domain of TIMP-2 is not required for the antimetalloproteinase activity but plays a major role in the high affinity cell surface binding of the inhibitor.
authors.affiliation: Childrens Hospital Los Angeles and the Department of Pediatrics, University of Southern California, Los Angeles 90027, USA.
authors.forename: Y C
authors.initials: YC
authors.lastname: Ko
authors.forename: K E
authors.initials: KE
authors.lastname: Langley
authors.forename: E A
authors.initials: EA
authors.lastname: Mendiaz
authors.forename: V P
authors.initials: VP
authors.lastname: Parker
authors.forename: S M
authors.initials: SM
authors.lastname: Taylor
authors.forename: Y A
authors.initials: YA
authors.lastname: DeClerck
mesh_terms: D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2
chemical_list: D018076:DNA, Complementary; D011506:Proteins; D019716:Tissue Inhibitor of Metalloproteinase-2; D008666:Metalloendopeptidases