Formation of reconstituted casein micelles with human beta-caseins and bovine kappa-casein.
Human beta-casein (CN) is the major protein of the human milk casein fraction (approximately 80%) and exists in six calcium-sensitive forms, having zero to five organic phosphates per molecule. The major forms are the doubly-phosphorylated (beta-CN-2P; approximately 30%) and the quadruply phosphorylated (beta-CN-4P; approximately 35%) forms. Although calcium-insensitive, kappa-CN is known for its role in preventing the precipitation of beta-CN in the presence of Ca+2, but it is not known how the different levels of phosphorylation may affect this. In the present investigation, turbidity, measured at 400 nm, was determined at increasing temperatures (4 up to 37 degrees C) for solutions of beta-CN-2P and beta-CN-4P (3 mg/ml in 0.02 M NaCl, 0.01 M imidazole, pH 7) individually and also mixed with bovine kappa-CN in 6/1 and 3/1 weight ratios of beta/kappa and containing 0, 5, and 10 mM Ca+2. The results indicate that the first step of micelle formation probably leads to polymers of limited size, the only complexes available to beta-CN-2P under most conditions. With beta-CN-4P, these polymers aggregate further to give reconstituted micelles, probably because of the ability to form crosslinks at this phosphorylation level. The formation of reconstituted micelles under various conditions of pH, Ca+2 concentration and kappa-CN content indicates that both hydrophobic interactions and Ca+2 bridges or crosslinks may contribute to protein aggregation and micelle building.
Sood
S M
SM
Department of Biochemistry, School of Medicine, Loma Linda University, CA 92350, USA. ssood@som.llu.edu
Erickson
G
G
Slattery
C W
CW
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008823:Micelles; D008892:Milk / Q000737:chemistry / Q000378:metabolism; D008895:Milk, Human / Q000737:chemistry* / Q000378:metabolism; D009391:Nephelometry and Turbidimetry; D010766:Phosphorylation; D011108:Polymers; D013696:Temperature
D002364:Caseins; D002614:Chelating Agents; D008823:Micelles; D011108:Polymers; D002118:Calcium