Abstract
Abstract Chromatography of bovine plasma albumin (BPA) on diethylaminoethyl cellulose resulted in a single asymmetric peak with considerable tailing. An identical result was obtained with half-cystinyl bovine plasma albumin (Cys-BPA), in which the single sulfhydryl group of BPA is in the form of a mixed disulfide with l-cysteine. This chromatographic behavior was related to the presence of albumins with different isoelectric points as revealed by isoelectric focusing of Cys-BPA on acrylamide gel containing 6 m urea. The two major components, about 76 and 20% of the sample, differed by 1 unit charge. This was shown by comparative isoelectric focusing studies of Cys-BPA and its mono-14C-acetylated derivative. The mono-14C-acetylated albumin and possibly the di- and triacetylated derivatives were prepared by the reaction of 14C-acetylsalicyclic acid with Cys-BPA. Investigation of chromatographic cuts, enriched with the different isoelectric components of Cys-BPA, indicated that these components were noninterconvertible on reversible denaturation or on treatment with thiol reagents, and that these components differed in their thermal and acid denaturation properties. The isoelectric heterogeneity of Cys-BPA is not a result of heterozygosity of the structural gene since studies of Cys-BPA isolated from six separate calves showed the same result. Also studies with albumins isolated from eight humans and four mice showed similar findings.
Highlights
Chromatographic and Isoelectric Heterogeneity of bovine plasma albumin (BPA)--In the present work most of the studies were made with crystalline BPA samples which had been isolated from pooled animals
Some of the studies were made with albumin isolated from the plasma of a single calf by
Prior to use the albumin samples were first converted to cystinyl bovine plasma albumin (Cys-BPA), freed of dimers and polymers by chromatography on Sephadex G-100 defatted with the acid-charcoal method [12]
Summary
Isolation of Albumin from Plasma of Single Donors-The blood immediately after collection was diluted with 0.11 part of a. Vol 246, No 1 solution of citric acid, sodium citrate, and dextrose (A.C.D. solution, Abbott). To the plasma was added saturated (NHJ&K& solution to give a final concentration of 55% in (NH&Sod. The precipitate was removed by centrifugation, and the supernatant was brought to 85% saturation by the addition of solid (KH4)2S04. The precipitated albumin was processed as described below
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