title: [Purification and characterization of the respiratory inhibitor RF from rabbit reticulocytes].
abstract: In the stroma-free hemolysate of rabbit reticulocytes there exists a respiratory inhibitor (RF) of protein nature. It could be purified by the following procedure: (NH4)2SO4-precipitation leads to DEAE-Sephadex A-50-chromatography leads to isoelectric focusing leads to gelfiltration on Sephadex G-200. In addition to the activity the identity of the inhibitor could be proved immunologically at all steps of the purification. A molecular weight of about 80,000 was determined by gelelectrophoresis in the SDS-mercaptoethanol-system. The protein has an IP of 5.55--5.65 and consists of one polypeptide chain. This could be shown both by SDS-gelelectrophoresis and by estimation of only one N-terminal amino acid residue (glycine). Only after oxidation with performic acid the amino acid analysis of the protein gives reproducible values. The sum of weight-% is about 80 (without Tyr). The polarity of the protein with 41 mol-% agrees with the polarity of soluble globular proteins investigated by other authors. Acid hydrolysis without preceeding performic acid oxidation gives a sum of amino acid residues of only 35--45 weight-%, the glycoprotein nature of the inhibitor could be shown by staining the SDS-gels with Schiff's reagents and by carbohydrate determination with anthrone reagent.
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011817:Rabbits; D012156:Reticulocytes / Q000032:analysis*
chemical_list: D001798:Blood Proteins
vernacular_title: Reinigung und Charakterisierung des Atmungshemmstoffes RF aus Kaninchenretikulozyten.