Structure, function and tissue forms of the C-terminal globular domain of collagen XVIII containing the angiogenesis inhibitor endostatin.
The C-terminal domain NC1 of mouse collagen XVIII (38 kDa) and the shorter mouse and human endostatins (22 kDa) were prepared in recombinant form from transfected mammalian cells. The NC1 domain aggregated non-covalently into a globular trimer which was partially cleaved by endogenous proteolysis into several monomers (25-32 kDa) related to endostatin. Endostatins were obtained in a highly soluble, monomeric form and showed a single N-terminal sequence which, together with other data, indicated a compact folding. Endostatins and NC1 showed a comparable binding activity for the microfibrillar fibulin-1 and fibulin-2, and for heparin. Domain NC1, however, was a distinctly stronger ligand than endostatin for sulfatides and the basement membrane proteins laminin-1 and perlecan. Immunological assays demonstrated endostatin epitopes on several tissue components (22-38 kDa) and in serum (120-300 ng/ml), the latter representing the smaller variants. The data indicated that the NC1 domain consists of an N-terminal association region (approximately 50 residues), a central protease-sensitive hinge region (approximately 70 residues) and a C-terminal stable endostatin domain (approximately 180 residues). They also demonstrated that proteolytic release of endostatin can occur through several pathways, which may lead to a switch from a matrix-associated to a more soluble endocrine form.
Max-Planck-Institut für Biochemie, Martinsried, Germany.
T
T
Sasaki
N
N
Fukai
K
K
Mann
W
W
Göhring
B R
BR
Olsen
R
R
Timpl
D000595:Amino Acid Sequence; D000818:Animals; D050260:Carbohydrate Metabolism; D002460:Cell Line; D003094:Collagen / Q000096:biosynthesis / Q000097:blood / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D043170:Collagen Type XVIII; D010450:Endopeptidases; D043169:Endostatins; D005822:Genetic Vectors; D006801:Humans; D006868:Hydrolysis; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D009928:Organ Specificity; D010446:Peptide Fragments / Q000096:biosynthesis / Q000097:blood / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
D043170:Collagen Type XVIII; D043169:Endostatins; D008024:Ligands; D010446:Peptide Fragments; D003094:Collagen; D010450:Endopeptidases