Determination of the disulfide bond pairings in human tissue factor pathway inhibitor purified from Escherichia coli.
The disulfide bond assignments of human alanyl tissue factor pathway inhibitor purified from Escherichia coli have been determined. This inhibitor of the extrinsic blood coagulation pathway possesses three Kunitz-type inhibitor domains, each containing three disulfide bonds. The disulfide bond pairings in domains 1 and 3 were determined by amino acid sequencing and mass spectrometry of peptides derived from a thermolysin digest. However, thermolysin digestion did not cleave any peptide bonds within domain 2. The disulfide bond pairings in domain 2 were determined by isolating it from the thermolysin treatment and subsequently cleaving it with pepsin and trypsin into peptides which yielded the three disulfide bond pairings in this domain. These results demonstrate that the disulfide pairings in each of the three domains of human tissue factor pathway inhibitor purified from Escherichia coli are homologous to each other and also to those in bovine pancreatic trypsin inhibitor.
Violand
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BN
Protiva Division, Monsanto Company, St. Louis, Missouri 63198, USA.
Schlittler
M R
MR
Duffin
K L
KL
Smith
C E
CE
D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000032:analysis* / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008074:Lipoproteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data
D000925:Anticoagulants; D004220:Disulfides; D008074:Lipoproteins; C051928:lipoprotein-associated coagulation inhibitor