Identification of human milk kappa-casein on polyacrylamide gels by differential staining with Ethyl-Stains-all and chymosin sensitivity.
Ethyl-Stains-all (ESA), a cationic carbocyanine dye that stains phosphorylated, sialylated, and unmodified proteins differentially, was used to stain a human casein fraction enriched for its kappa-casein-like characteristics. The staining properties and chymosin sensitivity of this fraction were compared with those of human milk and bovine casein proteins. Phosphorylated human and bovine beta caseins stained blue with ESA. The sialic acid-containing bovine kappa-casein stained blue-green. The human kappa-like fraction was enriched for a protein that stained blue-green with ESA. Both bovine kappa-casein and the human blue-green-staining protein were susceptible to chymosin digestion at lower concentrations of chymosin than that required for digestion of beta-caseins. In each case, following chymosin digestion, a green-staining peptide of lower molecular weight replaced the original protein and para-kappa-casein was formed. Identification of human kappa-casein on SDS-polyacrylamide gels was based on its differential staining with ESA and chymosin sensitivity with respect to beta-casein.
M R
MR
Green
D000818:Animals; D002232:Carbocyanines; D002364:Caseins / Q000032:analysis*; D002417:Cattle; D012085:Chymosin / Q000494:pharmacology*; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008895:Milk, Human / Q000032:analysis*; D011804:Quinolines; D013194:Staining and Labeling
D002232:Carbocyanines; D002364:Caseins; D004396:Coloring Agents; D011804:Quinolines; C017873:ethyl-stains-all; D012085:Chymosin