Molecular characterization of alpha-lactalbumin folding variants that induce apoptosis in tumor cells.
This study characterized a protein complex in human milk that induces apoptosis in tumor cells but spares healthy cells. The active fraction was purified from casein by anion exchange chromatography. Unlike other casein components the active fraction was retained by the ion exchanger and eluted after a high salt gradient. The active fraction showed N-terminal amino acid sequence identity with human milk alpha-lactalbumin and mass spectrometry ruled out post-translational modifications. Size exclusion chromatography resolved monomers and oligomers of alpha-lactalbumin that were characterized using UV absorbance, fluorescence, and circular dichroism spectroscopy. The high molecular weight oligomers were kinetically stable against dissociation into monomers and were found to have an essentially retained secondary structure but a less well organized tertiary structure. Comparison with native monomeric and molten globule alpha-lactalbumin showed that the active fraction contains oligomers of alpha-lactalbumin that have undergone a conformational switch toward a molten globule-like state. Oligomerization appears to conserve alpha-lactalbumin in a state with molten globule-like properties at physiological conditions. The results suggest differences in biological properties between folding variants of alpha-lactalbumin.
Svensson
M
M
Institute of Laboratory Medicine, Section of Microbiology, Immunology, and Glycobiology (MIG), Lund University, Sölvegatan 23, S-223 62 Lund, Sweden. Malin.Svensson@mig.lu.se
Sabharwal
H
H
Håkansson
A
A
Mossberg
A K
AK
Lipniunas
P
P
Leffler
H
H
Svanborg
C
C
Linse
S
S
D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects*; D019281:Dimerization; D005260:Female; D006801:Humans; D007768:Lactalbumin / Q000737:chemistry* / Q000494:pharmacology; D008895:Milk, Human / Q000737:chemistry*; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000473:pathology; D017510:Protein Folding; D013329:Structure-Activity Relationship
D007768:Lactalbumin