Processing and expression of rat and human clotting factor-X-encoding cDNAs.
The cDNA encoding clotting factor X, which participates in the middle stage of the blood coagulation cascade was cloned from a rat liver cDNA library. Sequencing of the rat factor-X-encoding cDNA revealed that this vitamin-K-dependent protein has a dibasic Arg-Arg sequence at the propeptide cleavage site, as occurs in other vitamin-K-dependent proteins. Although the human and rat deduced amino acid sequences are remarkably similar (76% identical), they do significantly differ in that human factor-X contains a unique Thr-Arg sequence at the propeptide cleavage site [Fung et al., Proc. Natl. Acad. Sci. USA 82 (1985) 3591-3595], where a dibasic sequence would normally be expected. This specific site is the recognition motif for the endoprotease, furin, which is located in the Golgi apparatus. Both rat and human cDNAs expressed in Cos-1 cells resulted in secretion of a mixture of single- and two-chain forms of factor X. The two-chain forms were devoid of the propeptide and were produced at similar rates by the transfected cells. The efficient processing of human factor X, when compared to rat factor X, may indicate that an additional protease(s), which recognizes the Thr-Arg motif, may be involved in proteolytic processing of the human enzyme.
Stanton
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Department of Medicine, Wake Forest University Medical Center, Winston Salem, NC 27157, USA.
Ross
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Hutson
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Wallin
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005170:Factor X / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D045683:Furin; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D051381:Rats; D013381:Subtilisins / Q000378:metabolism
D018076:DNA, Complementary; D005170:Factor X; D013381:Subtilisins; D045683:Furin