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Oligomerization-dependent regulation of motility and morphogenesis by the collagen XVIII NC1/endostatin domain.

Kuo CJ, LaMontagne KR, Garcia-Cardeña G, Ackley BD, Kalman D, Park S, Christofferson R, Kamihara J, Ding YH, Lo KM, Gillies S, Folkman J, Mulligan RC, Javaherian K - J. Cell Biol. (2001)

Bottom Line: Here, the c18 NC1 domain functioned as a motility-inducing factor regulating the extracellular matrix (ECM)-dependent morphogenesis of endothelial and other cell types.This motogenic activity required ES domain oligomerization, was dependent on rac, cdc42, and mitogen-activated protein kinase, and exhibited functional distinction from the archetypal motogenic scatter factors hepatocyte growth factor and macrophage stimulatory protein.These data indicate that the collagen XVIII NC1 region encodes a motogen strictly requiring ES domain oligomerization and suggest a previously unsuspected mechanism for ECM regulation of motility and morphogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Children's Hospital, Harvard Medical School, Boston. Massachusetts 02115, USA. cjkuo@stanford.edu

ABSTRACT
Collagen XVIII (c18) is a triple helical endothelial/epithelial basement membrane protein whose noncollagenous (NC)1 region trimerizes a COOH-terminal endostatin (ES) domain conserved in vertebrates, Caenorhabditis elegans and Drosophila. Here, the c18 NC1 domain functioned as a motility-inducing factor regulating the extracellular matrix (ECM)-dependent morphogenesis of endothelial and other cell types. This motogenic activity required ES domain oligomerization, was dependent on rac, cdc42, and mitogen-activated protein kinase, and exhibited functional distinction from the archetypal motogenic scatter factors hepatocyte growth factor and macrophage stimulatory protein. The motility-inducing and mitogen-activated protein kinase-stimulating activities of c18 NC1 were blocked by its physiologic cleavage product ES monomer, consistent with a proteolysis-dependent negative feedback mechanism. These data indicate that the collagen XVIII NC1 region encodes a motogen strictly requiring ES domain oligomerization and suggest a previously unsuspected mechanism for ECM regulation of motility and morphogenesis.

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Oligomerized collagen XV ES domains do not exhibit motogenic activity. HUVECs were plated on Matrigel in the presence of mNC1(c18) (50 nM), mNC1(c15) (100 nM), or Fc-mES(c15) (250 nM) and photographed after 20 h. Identical results were obtained upon application of factors to preformed tubules.
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Figure 4: Oligomerized collagen XV ES domains do not exhibit motogenic activity. HUVECs were plated on Matrigel in the presence of mNC1(c18) (50 nM), mNC1(c15) (100 nM), or Fc-mES(c15) (250 nM) and photographed after 20 h. Identical results were obtained upon application of factors to preformed tubules.

Mentions: Collagen XVIII and collagen XV define the multiplexin subclass of collagens, characterized by multiple interruptions in the triple helical region and a globular COOH-terminal ES domain. The NC1 domains of both c18 and c15 trimerize (Sasaki et al. 2000), and the cognate ES domains exhibit 60% amino acid identity (Muragaki et al. 1994; Oh et al. 1994; Rehn and Pihlajaniemi 1994). Despite these similarities, the c18 and c15 NC1 domains differ strikingly in their abilities to associate with heparin and zinc, in affinity for matrix components exhibited by monomers versus trimers, and a shorter “hinge” region in the NC1 domain in c15 (Sasaki et al. 2000). We produced recombinant c15 NC1 domains from the conditioned medium of transfected 293T cells but observed complete lack of inhibition of HUVEC tube formation on Matrigel at 100 nM, the highest concentration tested (Fig. 4). Similarly, dimeric Fc fusion proteins containing Fc at the NH2 terminus and the collagen XV ES domain at the COOH terminus (Fc-ES[c15]) neither scattered endothelial tubules (Fig. 4) nor antagonized the effects of Fc-ES(c18) at 30-fold molar excess (data not shown). Under these conditions, the motogenic properties of ES domain oligomers are thus highly specific for c18 and not c15, in accordance with divergent structural and functional properties (Muragaki et al. 1994; Oh et al. 1994; Rehn and Pihlajaniemi 1994; Sasaki et al. 2000).


Oligomerization-dependent regulation of motility and morphogenesis by the collagen XVIII NC1/endostatin domain.

Kuo CJ, LaMontagne KR, Garcia-Cardeña G, Ackley BD, Kalman D, Park S, Christofferson R, Kamihara J, Ding YH, Lo KM, Gillies S, Folkman J, Mulligan RC, Javaherian K - J. Cell Biol. (2001)

Oligomerized collagen XV ES domains do not exhibit motogenic activity. HUVECs were plated on Matrigel in the presence of mNC1(c18) (50 nM), mNC1(c15) (100 nM), or Fc-mES(c15) (250 nM) and photographed after 20 h. Identical results were obtained upon application of factors to preformed tubules.
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Related In: Results  -  Collection

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Figure 4: Oligomerized collagen XV ES domains do not exhibit motogenic activity. HUVECs were plated on Matrigel in the presence of mNC1(c18) (50 nM), mNC1(c15) (100 nM), or Fc-mES(c15) (250 nM) and photographed after 20 h. Identical results were obtained upon application of factors to preformed tubules.
Mentions: Collagen XVIII and collagen XV define the multiplexin subclass of collagens, characterized by multiple interruptions in the triple helical region and a globular COOH-terminal ES domain. The NC1 domains of both c18 and c15 trimerize (Sasaki et al. 2000), and the cognate ES domains exhibit 60% amino acid identity (Muragaki et al. 1994; Oh et al. 1994; Rehn and Pihlajaniemi 1994). Despite these similarities, the c18 and c15 NC1 domains differ strikingly in their abilities to associate with heparin and zinc, in affinity for matrix components exhibited by monomers versus trimers, and a shorter “hinge” region in the NC1 domain in c15 (Sasaki et al. 2000). We produced recombinant c15 NC1 domains from the conditioned medium of transfected 293T cells but observed complete lack of inhibition of HUVEC tube formation on Matrigel at 100 nM, the highest concentration tested (Fig. 4). Similarly, dimeric Fc fusion proteins containing Fc at the NH2 terminus and the collagen XV ES domain at the COOH terminus (Fc-ES[c15]) neither scattered endothelial tubules (Fig. 4) nor antagonized the effects of Fc-ES(c18) at 30-fold molar excess (data not shown). Under these conditions, the motogenic properties of ES domain oligomers are thus highly specific for c18 and not c15, in accordance with divergent structural and functional properties (Muragaki et al. 1994; Oh et al. 1994; Rehn and Pihlajaniemi 1994; Sasaki et al. 2000).

Bottom Line: Here, the c18 NC1 domain functioned as a motility-inducing factor regulating the extracellular matrix (ECM)-dependent morphogenesis of endothelial and other cell types.This motogenic activity required ES domain oligomerization, was dependent on rac, cdc42, and mitogen-activated protein kinase, and exhibited functional distinction from the archetypal motogenic scatter factors hepatocyte growth factor and macrophage stimulatory protein.These data indicate that the collagen XVIII NC1 region encodes a motogen strictly requiring ES domain oligomerization and suggest a previously unsuspected mechanism for ECM regulation of motility and morphogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Children's Hospital, Harvard Medical School, Boston. Massachusetts 02115, USA. cjkuo@stanford.edu

ABSTRACT
Collagen XVIII (c18) is a triple helical endothelial/epithelial basement membrane protein whose noncollagenous (NC)1 region trimerizes a COOH-terminal endostatin (ES) domain conserved in vertebrates, Caenorhabditis elegans and Drosophila. Here, the c18 NC1 domain functioned as a motility-inducing factor regulating the extracellular matrix (ECM)-dependent morphogenesis of endothelial and other cell types. This motogenic activity required ES domain oligomerization, was dependent on rac, cdc42, and mitogen-activated protein kinase, and exhibited functional distinction from the archetypal motogenic scatter factors hepatocyte growth factor and macrophage stimulatory protein. The motility-inducing and mitogen-activated protein kinase-stimulating activities of c18 NC1 were blocked by its physiologic cleavage product ES monomer, consistent with a proteolysis-dependent negative feedback mechanism. These data indicate that the collagen XVIII NC1 region encodes a motogen strictly requiring ES domain oligomerization and suggest a previously unsuspected mechanism for ECM regulation of motility and morphogenesis.

Show MeSH
Related in: MedlinePlus