Characterization of a cDNA coding for human factor X.

S P Leytus; D W Chung; W Kisiel; K Kurachi; E W Davie

doi:10.1073/pnas.81.12.3699

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Characterization of a cDNA coding for human factor X.

S P Leytus·D W Chung·W Kisiel·K Kurachi·E W Davie

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Abstract

A lambda gt11 cDNA library containing DNA inserts prepared from human liver mRNA has been screened with an antibody to human factor X, a plasma protein participating in the middle phase of the blood coagulation cascade. Ten positive clones were isolated from 2 X 10(6) phage and plaque purified. The cDNA in the phage containing the largest insert has been sequenced and shown to code for human factor X. This cDNA insert contained 1137 base pairs coding for a portion of the light chain of the molecule, a connecting region, the heavy chain, a stop codon, a short 3' noncoding region, and a poly(A) tail. The sequence of A-T-T-A-A-A, which functions as a potential recognition site for polyadenylylation or processing, was present in the 3' end of the coding sequence and preceded the stop codon of TGA by 1 base pair and the poly(A) tail by 14 base pairs. The amino acid sequence deduced from the cDNA indicated that factor X is synthesized as a single-chain polypeptide containing the light and heavy chains connected by an Arg-Lys-Arg tripeptide. The single-chain molecule is then converted to the light and heavy chains by cleavage of two (or more) internal peptide bonds. In plasma, these two chains are linked together by a disulfide bond. The DNA sequence coding for the active site of human factor X showed a high degree of identity with prothrombin and factor IX, two other vitamin K-dependent serine proteases that participate in blood coagulation. These data along with the protein sequence data previously published for the light chain of human factor X establish the complete amino acid sequence for the mature protein present in plasma.

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Amino Acid Sequence Base Sequence DNA Factor X Humans Liver Macromolecular Substances Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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S P Leytus, D W Chung, W Kisiel, K Kurachi, E W Davie. 1984. Characterization of a cDNA coding for human factor X.. https://doi.org/10.1073/pnas.81.12.3699

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