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Role of the netrin-like domain of procollagen C-proteinase enhancer-1 in the control of metalloproteinase activity.

Bekhouche M, Kronenberg D, Vadon-Le Goff S, Bijakowski C, Lim NH, Font B, Kessler E, Colige A, Nagase H, Murphy G, Hulmes DJ, Moali C - J. Biol. Chem. (2010)

Bottom Line: The presence of a C-terminal NTR domain in procollagen C-proteinase enhancers (PCPEs), proteins that stimulate the activity of astacin-like tolloid proteinases, raises the possibility that this might also have inhibitory activity.Here we show that both long and short forms of the PCPE-1 NTR domain, the latter beginning at the N-terminal cysteine known to be critical for TIMP activity, show no inhibition, at micromolar concentrations, of several members of the metzincin superfamily, including matrix metalloproteinase-2, bone morphogenetic protein-1 (a tolloid proteinase), and different ADAMTS (a disintegrin and a metalloproteinase with thrombospondin motifs) proteinases from the adamalysin family.In contrast, we report that the NTR domain within PCPE-1 leads to superstimulation of bone morphogenetic protein-1 activity in the presence of heparin and heparan sulfate.

View Article: PubMed Central - PubMed

Affiliation: From the Institut de Biologie et Chimie des Protéines, CNRS/Université de Lyon UMR 5086, IFR128, 69367 Lyon, France.

ABSTRACT
The netrin-like (NTR) domain is a feature of several extracellular proteins, most notably the N-terminal domain of tissue inhibitors of metalloproteinases (TIMPs), where it functions as a strong inhibitor of matrix metalloproteinases and some other members of the metzincin superfamily. The presence of a C-terminal NTR domain in procollagen C-proteinase enhancers (PCPEs), proteins that stimulate the activity of astacin-like tolloid proteinases, raises the possibility that this might also have inhibitory activity. Here we show that both long and short forms of the PCPE-1 NTR domain, the latter beginning at the N-terminal cysteine known to be critical for TIMP activity, show no inhibition, at micromolar concentrations, of several members of the metzincin superfamily, including matrix metalloproteinase-2, bone morphogenetic protein-1 (a tolloid proteinase), and different ADAMTS (a disintegrin and a metalloproteinase with thrombospondin motifs) proteinases from the adamalysin family. In contrast, we report that the NTR domain within PCPE-1 leads to superstimulation of bone morphogenetic protein-1 activity in the presence of heparin and heparan sulfate. These observations point to a new mechanism whereby binding to cell surface-associated or extracellular heparin-like sulfated glycosaminoglycans might provide a means to accelerate procollagen processing in specific cellular and extracellular microenvironments.

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Possible mechanism by which heparin-like glycosaminoglycans might superstimulate the enhancing activity of PCPE-1 in the C-terminal processing of procollagen III. PCPE-1 binds to procollagen III in the region of the C-propeptides with nanomolar affinity. This complex then binds to cell surface or extracellular matrix associated heparin-like glycosaminoglycans via interactions with PCPE-1 NTR and with procollagen III. In addition, BMP-1 interacts with PCPE-1 NTR, the CUB2-NTR linker, and heparin. These interactions might increase the local concentrations of the reactants and/or create a new recognition site that further facilitates the action of BMP-1, resulting in superstimulation. Approximate molecular dimensions are based on known low resolution structures (27, 64, 65). C1C2, CUB1CUB2.
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Figure 8: Possible mechanism by which heparin-like glycosaminoglycans might superstimulate the enhancing activity of PCPE-1 in the C-terminal processing of procollagen III. PCPE-1 binds to procollagen III in the region of the C-propeptides with nanomolar affinity. This complex then binds to cell surface or extracellular matrix associated heparin-like glycosaminoglycans via interactions with PCPE-1 NTR and with procollagen III. In addition, BMP-1 interacts with PCPE-1 NTR, the CUB2-NTR linker, and heparin. These interactions might increase the local concentrations of the reactants and/or create a new recognition site that further facilitates the action of BMP-1, resulting in superstimulation. Approximate molecular dimensions are based on known low resolution structures (27, 64, 65). C1C2, CUB1CUB2.

Mentions: PCPE-1 is known to bind to heparin, via its NTR domain (23, 41), with nanomolar affinity (41). Here we confirm this observation by surface plasmon resonance (Fig. 7) and show that although binding of heparin to mini-procollagen III is also relatively tight, binding of BMP-1 to heparin is much weaker. In contrast, the isolated C-propeptide trimer of procollagen III did not bind to heparin, indicating that the heparin-binding site is in the triple-helical or telopeptide region. By analogy with the known heparin binding sequence GIKGHR found near the C terminus of the collagen α1(I) chain triple-helical region (50), we propose that the site of interaction of heparin with procollagen III is an identical sequence at the same position (residues 1104–1109, numbered from the start of the signal sequence), which also occurs in mini-procollagen III. In addition, PCPE-1 has previously been shown to bind to mini-procollagen III, involving the CUB1CUB2 and C-propeptide regions, with nanomolar affinity (26), forming a 1:1 complex (23). Finally, here we show that PCPE-1 binds to BMP-1 mainly via its NTR domain, with micromolar affinity. Taken together, these data point to the possible formation of a complex involving all of the reactants, as depicted in Fig. 8.


Role of the netrin-like domain of procollagen C-proteinase enhancer-1 in the control of metalloproteinase activity.

Bekhouche M, Kronenberg D, Vadon-Le Goff S, Bijakowski C, Lim NH, Font B, Kessler E, Colige A, Nagase H, Murphy G, Hulmes DJ, Moali C - J. Biol. Chem. (2010)

Possible mechanism by which heparin-like glycosaminoglycans might superstimulate the enhancing activity of PCPE-1 in the C-terminal processing of procollagen III. PCPE-1 binds to procollagen III in the region of the C-propeptides with nanomolar affinity. This complex then binds to cell surface or extracellular matrix associated heparin-like glycosaminoglycans via interactions with PCPE-1 NTR and with procollagen III. In addition, BMP-1 interacts with PCPE-1 NTR, the CUB2-NTR linker, and heparin. These interactions might increase the local concentrations of the reactants and/or create a new recognition site that further facilitates the action of BMP-1, resulting in superstimulation. Approximate molecular dimensions are based on known low resolution structures (27, 64, 65). C1C2, CUB1CUB2.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2871463&req=5

Figure 8: Possible mechanism by which heparin-like glycosaminoglycans might superstimulate the enhancing activity of PCPE-1 in the C-terminal processing of procollagen III. PCPE-1 binds to procollagen III in the region of the C-propeptides with nanomolar affinity. This complex then binds to cell surface or extracellular matrix associated heparin-like glycosaminoglycans via interactions with PCPE-1 NTR and with procollagen III. In addition, BMP-1 interacts with PCPE-1 NTR, the CUB2-NTR linker, and heparin. These interactions might increase the local concentrations of the reactants and/or create a new recognition site that further facilitates the action of BMP-1, resulting in superstimulation. Approximate molecular dimensions are based on known low resolution structures (27, 64, 65). C1C2, CUB1CUB2.
Mentions: PCPE-1 is known to bind to heparin, via its NTR domain (23, 41), with nanomolar affinity (41). Here we confirm this observation by surface plasmon resonance (Fig. 7) and show that although binding of heparin to mini-procollagen III is also relatively tight, binding of BMP-1 to heparin is much weaker. In contrast, the isolated C-propeptide trimer of procollagen III did not bind to heparin, indicating that the heparin-binding site is in the triple-helical or telopeptide region. By analogy with the known heparin binding sequence GIKGHR found near the C terminus of the collagen α1(I) chain triple-helical region (50), we propose that the site of interaction of heparin with procollagen III is an identical sequence at the same position (residues 1104–1109, numbered from the start of the signal sequence), which also occurs in mini-procollagen III. In addition, PCPE-1 has previously been shown to bind to mini-procollagen III, involving the CUB1CUB2 and C-propeptide regions, with nanomolar affinity (26), forming a 1:1 complex (23). Finally, here we show that PCPE-1 binds to BMP-1 mainly via its NTR domain, with micromolar affinity. Taken together, these data point to the possible formation of a complex involving all of the reactants, as depicted in Fig. 8.

Bottom Line: The presence of a C-terminal NTR domain in procollagen C-proteinase enhancers (PCPEs), proteins that stimulate the activity of astacin-like tolloid proteinases, raises the possibility that this might also have inhibitory activity.Here we show that both long and short forms of the PCPE-1 NTR domain, the latter beginning at the N-terminal cysteine known to be critical for TIMP activity, show no inhibition, at micromolar concentrations, of several members of the metzincin superfamily, including matrix metalloproteinase-2, bone morphogenetic protein-1 (a tolloid proteinase), and different ADAMTS (a disintegrin and a metalloproteinase with thrombospondin motifs) proteinases from the adamalysin family.In contrast, we report that the NTR domain within PCPE-1 leads to superstimulation of bone morphogenetic protein-1 activity in the presence of heparin and heparan sulfate.

View Article: PubMed Central - PubMed

Affiliation: From the Institut de Biologie et Chimie des Protéines, CNRS/Université de Lyon UMR 5086, IFR128, 69367 Lyon, France.

ABSTRACT
The netrin-like (NTR) domain is a feature of several extracellular proteins, most notably the N-terminal domain of tissue inhibitors of metalloproteinases (TIMPs), where it functions as a strong inhibitor of matrix metalloproteinases and some other members of the metzincin superfamily. The presence of a C-terminal NTR domain in procollagen C-proteinase enhancers (PCPEs), proteins that stimulate the activity of astacin-like tolloid proteinases, raises the possibility that this might also have inhibitory activity. Here we show that both long and short forms of the PCPE-1 NTR domain, the latter beginning at the N-terminal cysteine known to be critical for TIMP activity, show no inhibition, at micromolar concentrations, of several members of the metzincin superfamily, including matrix metalloproteinase-2, bone morphogenetic protein-1 (a tolloid proteinase), and different ADAMTS (a disintegrin and a metalloproteinase with thrombospondin motifs) proteinases from the adamalysin family. In contrast, we report that the NTR domain within PCPE-1 leads to superstimulation of bone morphogenetic protein-1 activity in the presence of heparin and heparan sulfate. These observations point to a new mechanism whereby binding to cell surface-associated or extracellular heparin-like sulfated glycosaminoglycans might provide a means to accelerate procollagen processing in specific cellular and extracellular microenvironments.

Show MeSH