Hamster antithrombin III: purification, characterization and acute phase response.
Antithrombin III was purified to homogeneity from hamster plasma by affinity chromatography on heparin-agrose, ion-exchange chromatography on Mono Q and size-exclusion chromatography on TSK G3000SWG column with 50% yield. The molecular mass of hamster antithrombin III was estimated at 62.5 kDa and the absorption coefficient (A280 nm 1%, 1 cm) at 6.48 (in 0.1 M sodium phosphate pH 7.0). Several isoforms of the inhibitor were detected with the pI in range of 4.95-5.25. The protein contains all residues characteristic for complex-type carbohydrate chains. The N-terminal amino acid sequence shows 84% of identity to mouse and 76% to human analogue. The hamster antithrombin III gives low immunological cross-reactivity with antibodies to human antithrombin III. Initiation of the acute phase response only slightly affected the plasma concentration of inhibitor (+/- 10% within 72-h period). The kinetic data suggest high efficiency in bovine and human thrombin inhibition. In summary, the study shows only similarities between hamster and other mammal antithrombins.
Institute of Molecular Biology, Jagiellonian University, Kraków, Poland.
P
P
Mak
J J
JJ
Enghild
A
A
Dubin
D000210:Acute-Phase Reaction / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D000990:Antithrombin III / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D006224:Cricetinae; D006801:Humans; D007525:Isoelectric Focusing; D008647:Mesocricetus; D051379:Mice; D008969:Molecular Sequence Data; D012756:Sheep
D000990:Antithrombin III