Expression of Human Milk β-Casein in Escherichia coli: Comparison of Recombinant Protein with Native Isoforms

L. Hansson, S. Bergstrom, O. Hernell, B. Lonnerdal, A. K. Nilsson, M. Stromqvist

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Studies on physiological function and on structure-function relationships of human milk β-casein have been limited. In this study, we have introduced the human β-casein cDNA into vectors designed for expression in Escherichia coli. The inducible T7-based expression system resulted in high-level expression of recombinant β-casein. The recombinant β-casein, localized intracellularly in E. coli, was purified to homogeneity and compared with purified native β-casein, in particular with respect to phosphorylation. The E. coli-produced β-casein was found to comigrate with the full-length, nonphosphorylated native human β-casein iso-form on SDS-PAGE. An N-terminal peptide containing all tentative phosphorylation sites was isolated from the recombinant protein and analyzed by mass spectrometry. The molecular mass as well as the migration of this peptide on reversed-phase chromatography confirmed that it was unphosphorylated.

Original languageEnglish (US)
Pages (from-to)373-381
Number of pages9
JournalProtein Expression and Purification
Volume4
Issue number5
DOIs
StatePublished - Oct 1993

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Human Milk
Caseins
Recombinant Proteins
Escherichia coli
Protein Isoforms
Phosphorylation
Peptides
Molecular mass
Reverse-Phase Chromatography
Chromatography
Population Groups
Mass spectrometry
Polyacrylamide Gel Electrophoresis
Mass Spectrometry
Complementary DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression of Human Milk β-Casein in Escherichia coli : Comparison of Recombinant Protein with Native Isoforms. / Hansson, L.; Bergstrom, S.; Hernell, O.; Lonnerdal, B.; Nilsson, A. K.; Stromqvist, M.

In: Protein Expression and Purification, Vol. 4, No. 5, 10.1993, p. 373-381.

Research output: Contribution to journalArticle

Hansson, L. ; Bergstrom, S. ; Hernell, O. ; Lonnerdal, B. ; Nilsson, A. K. ; Stromqvist, M. / Expression of Human Milk β-Casein in Escherichia coli : Comparison of Recombinant Protein with Native Isoforms. In: Protein Expression and Purification. 1993 ; Vol. 4, No. 5. pp. 373-381.
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