Abstract
Abstract Bovine, porcine, and avian M-1 and bovine and porcine M-2 glycoproteins were isolated from blood plasmas by ammonium sulfate fractionation and chromatography on carboxymethyl cellulose columns. The glycoproteins were homogeneous when subjected to starch gel and acrylamide gel electrophoresis with a number of buffers from pH 4.5 to pH 8.7. When subjected to horizontal starch gel or horizontal acrylamide gel electrophoresis at pH 2.68, the procine and avian M-1 glycoproteins did not exhibit polymorphism, whereas the bovine M-1 glycoprotein was resolved into two zones in the starch gel and into four zones in the acrylamide gel electrophoresis. The sedimentation coefficients, s020,w, of the bovine, porcine, and avian M-1 glycoproteins were 2.8, 3.0, and 2.9, respectively. The molecular weights of the bovine, porcine, and avian M-1 glycoproteins and of the bovine and porcine M-2 glycoproteins were 42,000, 47,000, 44,000, 55,000, and 54,000, respectively. The glycoproteins were analyzed for hexosamine, hexose, sialic acid, fucose, amino acids, and nitrogen. The porcine M-1 and M-2 glycoproteins contained 25.3% and 14.8% fucose, respectively, while the bovine and avian glycoproteins contained less than 1% fucose. The amino acid composition of avian M-1 glycoprotein was especially different from the bovine and porcine M-1 glycoproteins in lysine, aspartic acid, isoleucine, tyrosine, and histidine contents. The bovine and porcine M-1 glycoproteins differed in their contents of arginine, threonine, serine, cystine, valine, leucine, and phenylalanine, whereas the M-2 glycoproteins differed in their contents of aspartic acid, threonine, glutamic acid, proline, cystine, valine, and leucine.
Highlights
M-2 glycoproteins were isolated from blood plasmas by ammonium sulfate fractionation and chromatography on carboxymethyl cellulose columns
For the determination of the amino acids, the glycoproteins were hydrolyzed in evacuated, sealed tubes for 24 and 72 hours with 5.7 N HCI at 105”
The properties of the isolated mobilities greater than (M-l) and M-2 bovine glycoproteins were similar to those reported by Bezkorovainy and Doherty [9, 10]
Summary
Bovine (domestic cattle), porcine (domestic swine), and avian (domestic fowl) bloods were collected in 0.1 volume of a 5 y0 solution of tetrasodium ethylenediaminetetraacetate in 0.9 yU aqueousNaCl, and the mixtures were centrifuged (2500 rpm) at 4”. For the determination of hexosamine [37] and hexose [38], for the estimation of the ratio of mannose to galactose, and for detection of hexoses by paper chromatography, t,he glycoproteins (approximately 10 mg) were hydrolyzed with a suspension (4 ml) of sulfonated polystyrene resin (Dowex 5OWX8, 200 to 400 mesh) in 0.075 N HCl (1:2, w/v) for 36 hours at 100” in a special oven with a rotating vertical disc [39]. For the detection of fucose by paper chromatography, the glycoproteins (2 to 3 mg) were hydrolyzed either with the above procedure (but with time of heating reduced to 12 hours) or with 0.1 N HzS04 (2 ml) for 2 hours at 100”. The fractions eluted from the column with water were subjected to descending paper chromatography for the detection of sugars. The acid eluates obtained after the resin hydrolysis [39]
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