Association of the quadruply phosphorylated beta-casein from human milk with the nonphosphorylated form.
Human beta-casein (beta-CN) is phosphorylated at levels from zero (beta-CN-0P) to five (beta-CN-5P). The major constituent is the 4P form (approximately 35%), whereas the 0P form (approximately 5%) has been implicated in the formation of a framework upon which the forms with higher levels of phosphorylation may aggregate. At 4 degrees C in 0.01 M imidazole and 0.02 M NaCl, pH 7, with a 1:1 (wt:wt) 0P:4P ratio and a total protein concentration of 3 mg/ml, the s20,w was 1.4 S (monomer). Laser light scattering gave a radius of approximately 4.5 nm. As the temperature, T, increased, s20,w increased to 2 S. At 25 degrees C, peaks of 9.5 S and 2 S were observed. This transition T was different from that of either form. At 37 degrees C, a single peak was again observed with s20,w of 17.5 S, compared with 42 S for the 0P and 14 S for the 4P form. Laser light scattering at 37 degrees C revealed a polymer of approximately 16 nm radius and D20,w of 1.55 cm2/s. A combination of D20,w and S20,w gave a relative molecular mass suggesting about 45 monomers per polymer. An incubation of 3 h or more at 37 degrees C caused further aggregation, characteristic of the 0P form, and supported the concept of framework formation. At pH 6.6, s20,w was 38 S compared with 1.4 S at pH 10.4. Hydrostatic pressure did not have a large effect but supported a soap micelle-like structure for the polymer. The turbidity of the mixture increased with the amount of CaCl2 and T until the protein precipitated. The properties of the 1:1 mixture of these human beta-CN are intermediate but probably more biased toward those for the 4P form.
Sood
S M
SM
Department of Biochemistry, School of Medicine, Loma Linda University, CA 92350, USA. ssood@som.llu.edu
Slattery
C W
CW
D002118:Calcium / Q000378:metabolism; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D008823:Micelles; D008895:Milk, Human / Q000737:chemistry* / Q000378:metabolism; D008970:Molecular Weight; D010766:Phosphorylation; D011312:Pressure; D012542:Scattering, Radiation; D013696:Temperature
D002364:Caseins; D008823:Micelles; D002118:Calcium