title: Collagenolytic Matrix Metalloproteinase Activities toward Peptomeric Triple-Helical Substrates.
abstract: Although collagenolytic matrix metalloproteinases (MMPs) possess common domain organizations, there are subtle differences in their processing of collagenous triple-helical substrates. In this study, we have incorporated peptoid residues into collagen model triple-helical peptides and examined MMP activities toward these peptomeric chimeras. Several different peptoid residues were incorporated into triple-helical substrates at subsites P3, P1, P1', and P10' individually or in combination, and the effects of the peptoid residues were evaluated on the activities of full-length MMP-1, MMP-8, MMP-13, and MMP-14/MT1-MMP. Most peptomers showed little discrimination between MMPs. However, a peptomer containing N-methyl Gly (sarcosine) in the P1' subsite and N-isobutyl Gly (NLeu) in the P10' subsite was hydrolyzed efficiently only by MMP-13 [nomenclature relative to the α1(I)772-786 sequence]. Cleavage site analysis showed hydrolysis at the Gly-Gln bond, indicating a shifted binding of the triple helix compared to the parent sequence. Favorable hydrolysis by MMP-13 was not due to sequence specificity or instability of the substrate triple helix but rather was based on the specific interactions of the P7' peptoid residue with the MMP-13 hemopexin-like domain. A fluorescence resonance energy transfer triple-helical peptomer was constructed and found to be readily processed by MMP-13, not cleaved by MMP-1 and MMP-8, and weakly hydrolyzed by MT1-MMP. The influence of the triple-helical structure containing peptoid residues on the interaction between MMP subsites and individual substrate residues may provide additional information about the mechanism of collagenolysis, the understanding of collagen specificity, and the design of selective MMP probes.
authors.affiliation: †Florida Atlantic University, 5353 Parkside Drive, Jupiter, Florida 33458, United States.|‡Torrey Pines Institute for Molecular Studies, 11350 Southwest Village Parkway, Port St. Lucie, Florida 34987, United States.
authors.forename: Maciej J
authors.initials: MJ
authors.lastname: Stawikowski
authors.affiliation: †Florida Atlantic University, 5353 Parkside Drive, Jupiter, Florida 33458, United States.|‡Torrey Pines Institute for Molecular Studies, 11350 Southwest Village Parkway, Port St. Lucie, Florida 34987, United States.
authors.forename: Roma
authors.initials: R
authors.lastname: Stawikowska
authors.affiliation: †Florida Atlantic University, 5353 Parkside Drive, Jupiter, Florida 33458, United States.|‡Torrey Pines Institute for Molecular Studies, 11350 Southwest Village Parkway, Port St. Lucie, Florida 34987, United States.|§The Scripps Research Institute/Scripps Florida, 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Gregg B
authors.initials: GB
authors.lastname: Fields
mesh_terms: D020781:Matrix Metalloproteinase 1 / Q000737:chemistry / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000737:chemistry / Q000378:metabolism; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry / Q000378:metabolism; D020784:Matrix Metalloproteinase 8 / Q000737:chemistry / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases; D020784:Matrix Metalloproteinase 8; D020781:Matrix Metalloproteinase 1; D053511:Matrix Metalloproteinase 14