Casein phosphopeptides influence calcium uptake by cultured human intestinal HT-29 tumor cells.
We investigated the direct effects of casein phosphopeptides (CPP), which are formed by the proteolytic degradation of alpha- and beta-caseins, on calcium uptake by human HT-29 intestinal tumor cells, which undergo an enterocytically oriented differentiation in culture. A commercial preparation containing a mixture of purified CPP and an individual CPP of 25 amino acids, both containing the characteristic Ca(2+) binding motif, ser(P)-ser(P)-ser(P)-glu-glu, were employed. The study was performed at the single-cell level and on a cell population and measured the changes in cytosolic calcium concentration before and after CPP addition. In the presence of 2 mmol/L extracellular calcium, both CPP preparations induced a transient rise of free intracellular calcium ions, which did not influence ATP-induced release of calcium from intracellular stores, and which disappeared completely in the absence of extracellular calcium. Pretreatment of these cells with thapsigargin, which completely empties the intracellular calcium stores, did not abolish the cell responses to CPP. Repetitive stimulation of HT-29 cells with CPP always elicited a transient calcium rise, suggesting a lack of desensitization. The CPP-stimulated cytosolic calcium rise was dependent on CPP dose, in a seemingly nonsaturating mode, and on cell numbers. All of this is consistent with the hypothesis that CPP do not influence membrane-bound receptors or ion channels, but may act as calcium ionophores or calcium carriers across the membrane. The reported findings provide a new basis on which to assess the possibility that CPP enhance calcium absorption and bioavailability in animals.
Ferraretto
A
A
Department of Medical Chemistry and Biochemistry, The Medical Faculty, University of Milan, L.I.T.A., 20090 Segrate, Milan, Italy.
Signorile
A
A
Gravaghi
C
C
Fiorilli
A
A
Tettamanti
G
G
D002118:Calcium / Q000378:metabolism*; D002364:Caseins / Q000494:pharmacology*; D002413:Cations, Divalent; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D005456:Fluorescent Dyes; D016257:Fura-2; D019073:HT29 Cells; D006801:Humans; D010748:Phosphopeptides / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence
D002364:Caseins; D002413:Cations, Divalent; D005456:Fluorescent Dyes; D010748:Phosphopeptides; D002118:Calcium; D016257:Fura-2