Rapid and Simple Procedure for Purifying PAS-4 Glycoprotein from Bovine Milk Fat Globule Membrane
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概要
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The isolation and partial characterization of PAS-4 glycoprotein (78kDa) from bovine milk fat globule membrane (MFGM) is described. PAS-4 was selectively extracted with Triton X-114 nonionic detergent and then fractionated on DEAE-Sepharose at pH 7. 5. The PAS-4 fraction that was not bound on DEAE-Sepharose gave a single band by SDS-PAGE. The recovery of PAS-4 was 57.4% from MFGM. An amino acid analysis found a high percentage of nonpolar residues. Approximately 7.2% of carbohydrate from PAS-4 was composed of mannose, galactose (Gal), N-acetylglucosamine, N-acetylgalactosamine (GalNAc), and sialic acid, most of the Gal and GalNAc in PAS-4 being released after mild alkaline hydrolysis. This indicated that PAS-4 contained both N- and O-linked sugar chains in concordance with the results of lectin affinity. PAS-4 had apparent isoelectric points of 7.45, 7.41, and 7.32, but these were shifted to pI 7.47 by a neuraminidase treatment. The apparent molecular weight of PAS-4 after de-glycosylation with N-glycanase was approximately 57,000 by SDS-PAGE.
- 社団法人日本農芸化学会の論文
- 1995-05-23
著者
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Azuma Norihiro
Department of Bioproductive Science, Faculty of Agriculture, Utsunomiya University
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Hwangbo Sik
Department Of Applied Biochemistry College Of Agriculture Utsunomiya University
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Azuma N
Department Of Biological Resource Chemistry Kyoto Prefectural University
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Azuma Norihiro
Department Of Agricultural Chemistry The University Of Tokyo
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KANNO Choemon
Department of Animal Science, Utsunomiya University
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Kanno C
Department Of Applied Biochemistry Utsunomiya University
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Kanno Choemon
Department Of Animal Science Utsunomiya University
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