Bothrops jararaca antithrombin: isolation, characterization and comparison with other animal antithrombins.
Antithrombin was purified from Bothrops jararaca plasma by affinity chromatography using HiTrap Heparin HP column, and its molecular weight, amino-terminal sequence, carbohydrate content, isoelectric point, inhibition of bovine thrombin, and immunological properties were studied and compared with previously described antithrombins. B. jararaca antithrombin is a single-chain glycoprotein with a total carbohydrate content of 18%. The molecular weight from SDS-PAGE was 61 kDa and the inhibitor exhibited an acidic isoelectric point (4.5). The amino-terminal sequence has been determined as His-Glu-Ser-Ser-Val-Gln-Asp-Ile-Ile-Thr, which is highly homologous to the terminal sequences of other animal antithrombins, indicating high amino acid conservation among several animals. Immunological cross-reactivity was observed among fish, frog, chicken, human, non-venomous snake and B. jararaca antithrombins. B. jararaca antithrombin showed inhibitory activity upon human and B. jararaca coagulation and amidolytic substrate S-2238.
de Morais
Karen Batista
KB
Laboratório de Fisiopatologia, Instituto Butantan, Avenida Vital Brazil, São Paulo, Brazil.
Vieira
Carolina Okamoto
CO
Hirata
Isaura Yoshico
IY
Tanaka-Azevedo
Anita Mitico
AM
D000595:Amino Acid Sequence; D000818:Animals; D000991:Antithrombins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D015153:Blotting, Western; D017837:Bothrops / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D013918:Thrombin Time
D000991:Antithrombins