Gelatin-binding region of human matrix metalloproteinase-2

Solution structure, dynamics, and function of the COL-23 two-domain construct

Klára Briknarová, Marion Gehrmann, L. Bányai, H. Tordai, László Pattha, Miguel Llinás

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Human matrix metalloproteinase-2 (MMP-2) contains an array of three fibronectin type II (FII) modules postulated to interact with gelatin (denatured collagen). Here, we verify that the NMR solution structure of the third FII repeat (COL-3) is similar to that of the second FII repeat (COL-2); characterize its ligand-binding properties; and derive dynamics properties and relative orientation in solution for the two domains of the COL-23 fragment, a construct comprising COL-2 and COL-3 in tandem, with each domain possessing a putative collagen-binding site. Interaction of the synthetic gelatin-like octadecapeptide (Pro-Pro-Gly)6 (PPG6) with COL-3 is weaker than with COL-2. We found that a synthetic peptide comprising segment 33-42 (peptide 33-42) from the MMP-2 prodomain interacts with COL-3 and, albeit with lower affinity, with COL-2 in a way that mimics PPG6 binding. COL-3 strongly prefers peptide 33-42 over PPG6, which suggests that intramolecular interactions with the prodomain could modulate binding of pro-MMP-2 to its gelatin substrate. In COL-23, the two modules retain their structural individuality and tumble independently. Overall, the NMR data indicate that the relative orientation of the modules in COL-23 is not fixed in solution, that the modules do not interact with one another, and that COL-23 is rather flexible. The binding sites face opposite each other, and their responses to, and normalized affinities for, the longer ligand PPG12 are virtually identical to those of the individual domains for PPG6, thus precluding cooperativity, although they may interact simultaneously with multiple sites of the extracellular matrix.

Original languageEnglish
Pages (from-to)27613-27621
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number29
DOIs
Publication statusPublished - Jul 20 2001

Fingerprint

prolyl-prolyl-glycine
Gelatin
Fibronectins
Matrix Metalloproteinase 2
Collagen
Binding Sites
Nuclear magnetic resonance
Ligands
Peptides
Individuality
Extracellular Matrix
tetracycline CMT-3
human MMP2 protein
poly(prolylprolylglycine)15
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Gelatin-binding region of human matrix metalloproteinase-2 : Solution structure, dynamics, and function of the COL-23 two-domain construct. / Briknarová, Klára; Gehrmann, Marion; Bányai, L.; Tordai, H.; Pattha, László; Llinás, Miguel.

In: Journal of Biological Chemistry, Vol. 276, No. 29, 20.07.2001, p. 27613-27621.

Research output: Contribution to journalArticle

@article{acf931c8bddb4153b3abbb6ca911b429,
title = "Gelatin-binding region of human matrix metalloproteinase-2: Solution structure, dynamics, and function of the COL-23 two-domain construct",
abstract = "Human matrix metalloproteinase-2 (MMP-2) contains an array of three fibronectin type II (FII) modules postulated to interact with gelatin (denatured collagen). Here, we verify that the NMR solution structure of the third FII repeat (COL-3) is similar to that of the second FII repeat (COL-2); characterize its ligand-binding properties; and derive dynamics properties and relative orientation in solution for the two domains of the COL-23 fragment, a construct comprising COL-2 and COL-3 in tandem, with each domain possessing a putative collagen-binding site. Interaction of the synthetic gelatin-like octadecapeptide (Pro-Pro-Gly)6 (PPG6) with COL-3 is weaker than with COL-2. We found that a synthetic peptide comprising segment 33-42 (peptide 33-42) from the MMP-2 prodomain interacts with COL-3 and, albeit with lower affinity, with COL-2 in a way that mimics PPG6 binding. COL-3 strongly prefers peptide 33-42 over PPG6, which suggests that intramolecular interactions with the prodomain could modulate binding of pro-MMP-2 to its gelatin substrate. In COL-23, the two modules retain their structural individuality and tumble independently. Overall, the NMR data indicate that the relative orientation of the modules in COL-23 is not fixed in solution, that the modules do not interact with one another, and that COL-23 is rather flexible. The binding sites face opposite each other, and their responses to, and normalized affinities for, the longer ligand PPG12 are virtually identical to those of the individual domains for PPG6, thus precluding cooperativity, although they may interact simultaneously with multiple sites of the extracellular matrix.",
author = "Kl{\'a}ra Briknarov{\'a} and Marion Gehrmann and L. B{\'a}nyai and H. Tordai and L{\'a}szl{\'o} Pattha and Miguel Llin{\'a}s",
year = "2001",
month = "7",
day = "20",
doi = "10.1074/jbc.M101105200",
language = "English",
volume = "276",
pages = "27613--27621",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "29",

}

TY - JOUR

T1 - Gelatin-binding region of human matrix metalloproteinase-2

T2 - Solution structure, dynamics, and function of the COL-23 two-domain construct

AU - Briknarová, Klára

AU - Gehrmann, Marion

AU - Bányai, L.

AU - Tordai, H.

AU - Pattha, László

AU - Llinás, Miguel

PY - 2001/7/20

Y1 - 2001/7/20

N2 - Human matrix metalloproteinase-2 (MMP-2) contains an array of three fibronectin type II (FII) modules postulated to interact with gelatin (denatured collagen). Here, we verify that the NMR solution structure of the third FII repeat (COL-3) is similar to that of the second FII repeat (COL-2); characterize its ligand-binding properties; and derive dynamics properties and relative orientation in solution for the two domains of the COL-23 fragment, a construct comprising COL-2 and COL-3 in tandem, with each domain possessing a putative collagen-binding site. Interaction of the synthetic gelatin-like octadecapeptide (Pro-Pro-Gly)6 (PPG6) with COL-3 is weaker than with COL-2. We found that a synthetic peptide comprising segment 33-42 (peptide 33-42) from the MMP-2 prodomain interacts with COL-3 and, albeit with lower affinity, with COL-2 in a way that mimics PPG6 binding. COL-3 strongly prefers peptide 33-42 over PPG6, which suggests that intramolecular interactions with the prodomain could modulate binding of pro-MMP-2 to its gelatin substrate. In COL-23, the two modules retain their structural individuality and tumble independently. Overall, the NMR data indicate that the relative orientation of the modules in COL-23 is not fixed in solution, that the modules do not interact with one another, and that COL-23 is rather flexible. The binding sites face opposite each other, and their responses to, and normalized affinities for, the longer ligand PPG12 are virtually identical to those of the individual domains for PPG6, thus precluding cooperativity, although they may interact simultaneously with multiple sites of the extracellular matrix.

AB - Human matrix metalloproteinase-2 (MMP-2) contains an array of three fibronectin type II (FII) modules postulated to interact with gelatin (denatured collagen). Here, we verify that the NMR solution structure of the third FII repeat (COL-3) is similar to that of the second FII repeat (COL-2); characterize its ligand-binding properties; and derive dynamics properties and relative orientation in solution for the two domains of the COL-23 fragment, a construct comprising COL-2 and COL-3 in tandem, with each domain possessing a putative collagen-binding site. Interaction of the synthetic gelatin-like octadecapeptide (Pro-Pro-Gly)6 (PPG6) with COL-3 is weaker than with COL-2. We found that a synthetic peptide comprising segment 33-42 (peptide 33-42) from the MMP-2 prodomain interacts with COL-3 and, albeit with lower affinity, with COL-2 in a way that mimics PPG6 binding. COL-3 strongly prefers peptide 33-42 over PPG6, which suggests that intramolecular interactions with the prodomain could modulate binding of pro-MMP-2 to its gelatin substrate. In COL-23, the two modules retain their structural individuality and tumble independently. Overall, the NMR data indicate that the relative orientation of the modules in COL-23 is not fixed in solution, that the modules do not interact with one another, and that COL-23 is rather flexible. The binding sites face opposite each other, and their responses to, and normalized affinities for, the longer ligand PPG12 are virtually identical to those of the individual domains for PPG6, thus precluding cooperativity, although they may interact simultaneously with multiple sites of the extracellular matrix.

UR - http://www.scopus.com/inward/record.url?scp=0035920182&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035920182&partnerID=8YFLogxK

U2 - 10.1074/jbc.M101105200

DO - 10.1074/jbc.M101105200

M3 - Article

VL - 276

SP - 27613

EP - 27621

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 29

ER -