title: Colloidal calcium phosphate in the reconstituted milk micelle may direct wild-type recombinant human beta-casein to fold like the native protein.
abstract: Native human beta-casein (CN) at all phosphorylation levels exhibits reproducible behavior and appears to have a unique, stable folding pattern. In contrast, the recombinant non-phosphorylated form of human beta-CN (beta-CN-0P) with the exact amino acid sequence (wild-type), expressed and purified from Escherichia coli, differs greatly in its behavior from the native protein and the complexes formed are unstable to thermal cycling. However, when it was incorporated into reconstituted milk micelles, using bovine kappa-CN at a kappa/beta molar ratio of 1/3 with added Ca2+ ions and inorganic phosphate (P(i)) at levels that would ordinarily precipitate, its association behavior vs. temperature as monitored by turbidity (OD(400 nm)) approximated that of native beta-CN-0P. This suggests that the milk micelle system, and particularly the colloidal calcium phosphate, may act as a 'molecular chaperon' to direct the folding of the molecule into the highly stable conformation found in the purified native human beta-CN molecule.
authors.affiliation: Department of Biochemistry and Microbiology, Biochemistry Division School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA. ssood@som.llu.edu
authors.forename: Satish M
authors.initials: SM
authors.lastname: Sood
authors.forename: Grant
authors.initials: G
authors.lastname: Erickson
authors.forename: Harbor
authors.initials: H
authors.lastname: Jhawar
authors.forename: Charles W
authors.initials: CW
authors.lastname: Slattery
mesh_terms: D000818:Animals; D002130:Calcium Phosphates / Q000737:chemistry*; D002364:Caseins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003102:Colloids; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008823:Micelles; D008892:Milk / Q000737:chemistry*; D009391:Nephelometry and Turbidimetry; D017510:Protein Folding; D011994:Recombinant Proteins; D013696:Temperature
chemical_list: D002130:Calcium Phosphates; D002364:Caseins; D003102:Colloids; D008823:Micelles; D011994:Recombinant Proteins; C020243:calcium phosphate