Casein-derived bioactive phosphopeptides

Role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells

Anita Ferraretto, Claudia Gravaghi, Amelia Fiorilli, Guido Tettamanti

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Casein phosphopeptides β-CN(1-25)4P and α s1-CN(59-79)5P, from β- and αs1-casein, respectively, both carrying the characteristic 'acidic motif' Ser(P)-Ser(P)-Ser(P)-Glu-Glu, were chemically synthesized and administered to HT-29 cells differentiated in culture, which are a used model of intestinal epithelium for absorption studies. Both casein phosphopeptides caused an increase of [Ca2+]i due to influx of extracellular Ca 2+. The response was quantitatively higher with β-CN(1-25)4P than αs1-CN(59-79)5P. The synthetic peptide corresponding to the 'acidic motif' was ineffective and the dephosphorylated form of β-CN(1-25)4P almost inactive. The lack of the N-terminally located five amino acids, or sequence modifications within the N-terminal segment of β-CN(1-25)4P, caused a total loss of activity, whereas the lack of the C-terminal segment preserved activity. In conclusion, the influx of calcium into HT-29 cells caused by β-CN(1-25)4P appears to depend on the phosphorylated 'acidic motif' and the preceding N-terminal region.

Original languageEnglish
Pages (from-to)92-98
Number of pages7
JournalFEBS Letters
Volume551
Issue number1-3
DOIs
Publication statusPublished - Sep 11 2003

Fingerprint

Phosphopeptides
HT29 Cells
Phosphorylation
Caseins
Tumors
Cells
Calcium
glutamyl-glutamic acid
Neoplasms
Intestinal Absorption
Intestinal Mucosa
Amino Acid Sequence
Amino Acids
Peptides

Keywords

  • Calcium imaging
  • Casein phosphopeptide
  • Fura-2
  • HT-29 cell

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Casein-derived bioactive phosphopeptides : Role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells. / Ferraretto, Anita; Gravaghi, Claudia; Fiorilli, Amelia; Tettamanti, Guido.

In: FEBS Letters, Vol. 551, No. 1-3, 11.09.2003, p. 92-98.

Research output: Contribution to journalArticle

Ferraretto, Anita ; Gravaghi, Claudia ; Fiorilli, Amelia ; Tettamanti, Guido. / Casein-derived bioactive phosphopeptides : Role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells. In: FEBS Letters. 2003 ; Vol. 551, No. 1-3. pp. 92-98.
@article{d749fecf784349a894cbcb3ed74f1ff1,
title = "Casein-derived bioactive phosphopeptides: Role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells",
abstract = "Casein phosphopeptides β-CN(1-25)4P and α s1-CN(59-79)5P, from β- and αs1-casein, respectively, both carrying the characteristic 'acidic motif' Ser(P)-Ser(P)-Ser(P)-Glu-Glu, were chemically synthesized and administered to HT-29 cells differentiated in culture, which are a used model of intestinal epithelium for absorption studies. Both casein phosphopeptides caused an increase of [Ca2+]i due to influx of extracellular Ca 2+. The response was quantitatively higher with β-CN(1-25)4P than αs1-CN(59-79)5P. The synthetic peptide corresponding to the 'acidic motif' was ineffective and the dephosphorylated form of β-CN(1-25)4P almost inactive. The lack of the N-terminally located five amino acids, or sequence modifications within the N-terminal segment of β-CN(1-25)4P, caused a total loss of activity, whereas the lack of the C-terminal segment preserved activity. In conclusion, the influx of calcium into HT-29 cells caused by β-CN(1-25)4P appears to depend on the phosphorylated 'acidic motif' and the preceding N-terminal region.",
keywords = "Calcium imaging, Casein phosphopeptide, Fura-2, HT-29 cell",
author = "Anita Ferraretto and Claudia Gravaghi and Amelia Fiorilli and Guido Tettamanti",
year = "2003",
month = "9",
day = "11",
doi = "10.1016/S0014-5793(03)00741-5",
language = "English",
volume = "551",
pages = "92--98",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1-3",

}

TY - JOUR

T1 - Casein-derived bioactive phosphopeptides

T2 - Role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells

AU - Ferraretto, Anita

AU - Gravaghi, Claudia

AU - Fiorilli, Amelia

AU - Tettamanti, Guido

PY - 2003/9/11

Y1 - 2003/9/11

N2 - Casein phosphopeptides β-CN(1-25)4P and α s1-CN(59-79)5P, from β- and αs1-casein, respectively, both carrying the characteristic 'acidic motif' Ser(P)-Ser(P)-Ser(P)-Glu-Glu, were chemically synthesized and administered to HT-29 cells differentiated in culture, which are a used model of intestinal epithelium for absorption studies. Both casein phosphopeptides caused an increase of [Ca2+]i due to influx of extracellular Ca 2+. The response was quantitatively higher with β-CN(1-25)4P than αs1-CN(59-79)5P. The synthetic peptide corresponding to the 'acidic motif' was ineffective and the dephosphorylated form of β-CN(1-25)4P almost inactive. The lack of the N-terminally located five amino acids, or sequence modifications within the N-terminal segment of β-CN(1-25)4P, caused a total loss of activity, whereas the lack of the C-terminal segment preserved activity. In conclusion, the influx of calcium into HT-29 cells caused by β-CN(1-25)4P appears to depend on the phosphorylated 'acidic motif' and the preceding N-terminal region.

AB - Casein phosphopeptides β-CN(1-25)4P and α s1-CN(59-79)5P, from β- and αs1-casein, respectively, both carrying the characteristic 'acidic motif' Ser(P)-Ser(P)-Ser(P)-Glu-Glu, were chemically synthesized and administered to HT-29 cells differentiated in culture, which are a used model of intestinal epithelium for absorption studies. Both casein phosphopeptides caused an increase of [Ca2+]i due to influx of extracellular Ca 2+. The response was quantitatively higher with β-CN(1-25)4P than αs1-CN(59-79)5P. The synthetic peptide corresponding to the 'acidic motif' was ineffective and the dephosphorylated form of β-CN(1-25)4P almost inactive. The lack of the N-terminally located five amino acids, or sequence modifications within the N-terminal segment of β-CN(1-25)4P, caused a total loss of activity, whereas the lack of the C-terminal segment preserved activity. In conclusion, the influx of calcium into HT-29 cells caused by β-CN(1-25)4P appears to depend on the phosphorylated 'acidic motif' and the preceding N-terminal region.

KW - Calcium imaging

KW - Casein phosphopeptide

KW - Fura-2

KW - HT-29 cell

UR - http://www.scopus.com/inward/record.url?scp=0142134260&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0142134260&partnerID=8YFLogxK

U2 - 10.1016/S0014-5793(03)00741-5

DO - 10.1016/S0014-5793(03)00741-5

M3 - Article

VL - 551

SP - 92

EP - 98

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-3

ER -