Structural studies on casein micelles of human milk: dissociation of beta-casein of different phosphorylation levels induced by cooling and ethylenediaminetetraacetate.
Information on the structure of human casein micelles has been obtained from dissociation of beta-casein (CN). Two approaches were used: cooling at 4 degrees C and addition of EDTA. An initial loss of about 80% of the protein optical density occurred upon cooling to 4 degrees C. Dissociation was time dependent, and at > or = 24 h about 10% remained. However, mean size and voluminosity of micelles increased, as indicated by laser light scattering and viscosity measurements. This process was reversible, and 95% of the protein reentered the micelles upon incubation for 3 h at 37 degrees C. Upon cooling, amounts of nonphosphorylated beta-CN increased, and singly phosphorylated beta-CN levels were almost constant relative to the total beta-CN in micelles. Upon addition of EDTA (0 to 5 mM), the forms with three to five phosphates were the major dissociating constituents; EDTA that was added by dialysis produced similar results but at lower concentrations. These data suggest that, in the absence of significant amounts of alpha s1-CN, nonphosphorylated and singly phosphorylated human beta-CN may form a framework, as proposed for alpha s1-CN for bovine milk, along with the colloidal calcium phosphate for the development of the final micelle structure by addition of the more highly phosphorylated forms. The results also indicate that human casein micelles have a less rigid structure than those of other species.
Sood
S M
SM
Department of Biochemistry, Loma Linda University School of Medicine, CA 92350, USA.
Herbert
P J
PJ
Slattery
C W
CW
D002118:Calcium / Q000032:analysis; D002364:Caseins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology*; D003080:Cold Temperature; D004492:Edetic Acid / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008823:Micelles; D008895:Milk, Human / Q000737:chemistry*; D010766:Phosphorylation; D013696:Temperature
D002364:Caseins; D002614:Chelating Agents; D008823:Micelles; D004492:Edetic Acid; D002118:Calcium