A novel human metalloprotease synthesized in the liver and secreted into the blood: possibly, the von Willebrand factor-cleaving protease?
We identified a novel metalloprotease, which could be responsible for cleaving the Tyr842-Met843 peptide bond of von Willebrand factor (vWF). This metalloprotease was purified from Cohn Fraction-I precipitate of human pooled plasma by the combination of gel filtration, DEAE chromatography, and preparative polyacrylamide gel electrophoresis in the presence of SDS. The NH2-terminal amino acid sequence of the isolated protein was: AAGGILHLELLVAVGPDVFQAHQEDTRRY. Based on this sequence, we searched human genomic and EST databases, and identified compatible nucleotide sequences. These results suggested that this protein is a novel metalloprotease, a member of the family of a disintegrin and metalloprotease with thrombospondin type-1 motifs (ADAMTS), and its genomic DNA was mapped to human chromosome 9q34. Multiple human tissue northern blotting analysis indicated that the mRNA encoding this protease spanned approximately 5 kilobases and was uniquely expressed in the liver. Furthermore, we determined the cDNA sequence encoding this protease, and found that this protease was comprised of a signal peptide, a proregion followed by the putative furin cleavage site, a reprolysin-type zinc-metalloprotease domain, a disintegrin-like domain, a thrombospondin type-1 (TSP1) motif, a cysteine-rich region, a spacer domain, and COOH-terminal TSP1 motif repeats.
Soejima
K
K
First Research Departmen, The Chemo-Sero-Therapeutic Research Institute, Kumamoto 869-1298, Japan. soejima@kaketsuken.or.jp
Mimura
N
N
Hirashima
M
M
Maeda
H
H
Hamamoto
T
T
Nakagaki
T
T
Nozaki
C
C
D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; D017398:Alternative Splicing; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D019483:Disintegrins / Q000737:chemistry; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000096:biosynthesis; D019700:Thrombospondin 1 / Q000737:chemistry; D014018:Tissue Distribution; D014158:Transcription, Genetic; D014841:von Willebrand Factor / Q000378:metabolism*
D019483:Disintegrins; D020033:Protein Isoforms; D012333:RNA, Messenger; D019700:Thrombospondin 1; D014841:von Willebrand Factor; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human