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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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Circular dichroism spectra of the PCPE-1 NTR domain. The spectra for NTRs and NTRt were measured at 25 °C, and that for NTRt was also measured at 65 °C.
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Figure 2: Circular dichroism spectra of the PCPE-1 NTR domain. The spectra for NTRs and NTRt were measured at 25 °C, and that for NTRt was also measured at 65 °C.

Mentions: By circular dichroism spectroscopy, NTRs and NTRt were found to have similar spectra in the 180–260-nm range (Fig. 2) and similar to that of recombinant PCPE-1 NTR previously used for structural studies (1, 10). In addition, this domain was found to be remarkably stable to temperature, with a CD spectrum that remained essentially unchanged in the temperature range 25–65 °C (Fig. 2). We conclude that both NTRs and NTRt were correctly folded.


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)

Circular dichroism spectra of the PCPE-1 NTR domain. The spectra for NTRs and NTRt were measured at 25 °C, and that for NTRt was also measured at 65 °C.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Circular dichroism spectra of the PCPE-1 NTR domain. The spectra for NTRs and NTRt were measured at 25 °C, and that for NTRt was also measured at 65 °C.
Mentions: By circular dichroism spectroscopy, NTRs and NTRt were found to have similar spectra in the 180–260-nm range (Fig. 2) and similar to that of recombinant PCPE-1 NTR previously used for structural studies (1, 10). In addition, this domain was found to be remarkably stable to temperature, with a CD spectrum that remained essentially unchanged in the temperature range 25–65 °C (Fig. 2). We conclude that both NTRs and NTRt were correctly folded.

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