Cloning of the full-length cDNA of porcine antithrombin III and comparison with its human homolog.
The characterization of porcine antithrombin III (ATIII)-a highly powerful anticoagulant-is essential for using porcine liver in xenotransplantation applications. The objective of this study was to clarify the functions of porcine ATIII through comparison with human ATIII. We cloned porcine ATIII and compared its important functional sites with those of human ATIII. The full-length cDNA of porcine ATIII was cloned by screening a porcine liver cDNA library, and the ATIII activities of 23 pigs were determined. The full-length cDNA of porcine ATIII spanned 1498 bp and encoded 463 amino acids. Porcine ATIII shared 87.67% nucleotide identity and 89.06% amino acid identity with human ATIII. Complete identity was found at active center Arg393-Ser394, and remarkably high similarities were found at 2 critical heparin-binding sites (residues 41 through 49 and 114 through 156) and in some key residues involved in heparin binding. An ATIII assay found no significant difference between porcine and human plasma. The high level of similarity between porcine ATIII and human ATIII suggests that porcine ATIII will function in a manner similar to human ATIII in xenotransplantation.
Chen
Younan
Y
Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, Sichuan University, China.
Tan
Weidong
W
Qin
Shengfang
S
Zhang
Jie
J
Bu
Hong
H
Li
Youping
Y
Lu
Yanrong
Y
Cheng
Jingqiu
J
D000595:Amino Acid Sequence; D000818:Animals; D000990:Antithrombin III / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013552:Swine
D018076:DNA, Complementary; D000990:Antithrombin III