Purification and characterization of acute phase rat plasma thiostatin.
Thiostatin was purified from acute phase plasma of turpentine-treated rats by a novel, single-step carboxymethyl-papain Sepharose 4B column chromatographic procedure. Purified thiostatin appeared as a single band in SDS-PAGE with an estimated molecular weight of 68,000. Western blot with polyclonal rabbit anti-thiostatin IgG confirmed a homogeneous immuno-reactive 68 kDa species. Specific activity, as determined by enzyme-linked immunosorbent assay (ELISA), was 0.972 mg kininogen equivalent per mg protein. The yield of thiostatin exceeded 60% and the protein was purified 10.7-fold.
Rusiniak
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Department of Biochemical Pharmacology, State University of New York at Buffalo, New York 14260.
Wagh
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Bedi
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D000209:Acute-Phase Proteins / Q000302:isolation & purification*; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose / Q000379:methods*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007704:Kininogens / Q000097:blood / Q000302:isolation & purification*; D008297:Male; D010206:Papain / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014425:Turpentine / Q000494:pharmacology
D000209:Acute-Phase Proteins; D015853:Cysteine Proteinase Inhibitors; D007704:Kininogens; D010206:Papain; D014425:Turpentine