NC1 domain of human type VIII collagen (alpha 1) inhibits bovine aortic endothelial cell proliferation and causes cell apoptosis.
Endostatin, a natural angiogenesis inhibitor, had been identified for years. It opened a new approach for cancer therapy. Sequence analysis revealed that endostatin is the NC1 domain (non-triple-helical domain) of collagen XVIII. In this report, the cDNA of NC1 domain of type VIII collagen (alpha 1) was cloned and expressed as soluble form in Escherichia coli. The recombinant protein was purified with Ni-NTA agarose column and named as vastatin. It inhibited the proliferation of bovine aortic endothelial (BAE) cell stimulated by basic fibroblast growth factor (bFGF) in a dose-dependent manner. The ED(50) of vastatin was 0.6 microg/ml, while the ED(50) of endostatin was 0.5 microg/ml. Treatment of BAE cell with vastatin caused G(0)-G(1) arrest and cell apoptosis. It is interesting that sequence analysis showed that there was only about 12% amino acid sequence homology between vastatin and endostatin. The structure-function relationship of these angiogenesis molecules remains to be elucidated.
Xu
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R
Institute of Biochemistry and Cell Biology, Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, People's Republic of China.
Yao
Z Y
ZY
Xin
L
L
Zhang
Q
Q
Li
T P
TP
Gan
R B
RB
D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002417:Cattle; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D024102:Collagen Type VIII / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D043170:Collagen Type XVIII; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D043169:Endostatins; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid
D020533:Angiogenesis Inhibitors; D024102:Collagen Type VIII; D043170:Collagen Type XVIII; D017931:DNA Primers; D018076:DNA, Complementary; D043169:Endostatins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C441971:vastatin; D003094:Collagen