Isolation of mitogenic glycophosphopeptides from cheese whey protein concentrate.
We investigated the immunological function of cheese whey protein concentrate (CWPC), which is a by-product of cheese production, using mitogenic activity in murine splenocytes as an index. A fraction isolated by gel filtration and anion exchange chromatography of CWPC showed high mitogenic activity, comparable to the activity of lipopolysaccharide (LPS). The fraction was detected as a single band on SDS-PAGE. It contained calcium, inorganic phosphorus, and carbohydrate, indicating the active component to be a glycophosphopeptide (GPP). Since Pronase digestion of GPP did not reduce its mitogenic activity, carbohydrate rather than peptide may be important in the activity. When applied on an anti-beta-caseinophosphopeptide (beta-CPP) antibody affinity column, the GPP was separated into two components, one with affinity to beta-CPP and the other without such affinity. Both the components contained N-linked oligosaccharide chains and had the mitogenic activity. These results demonstrate that cheese whey contains a GPP having strong mitogenic activity.
College of Agriculture and Veterinary Medicine, Nihon University, Tokyo, Japan.
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Yun
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Sugita-Konishi
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Kumagai
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Yamauchi
D000276:Adjuvants, Immunologic / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002611:Cheese / Q000032:analysis*; D006020:Glycopeptides / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008894:Milk Proteins / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D008934:Mitogens / Q000302:isolation & purification* / Q000494:pharmacology*; D010748:Phosphopeptides / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology*; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D000067816:Whey Proteins
D000276:Adjuvants, Immunologic; D002241:Carbohydrates; D006020:Glycopeptides; D008894:Milk Proteins; D008934:Mitogens; D010748:Phosphopeptides; D000067816:Whey Proteins