Yuriko Adkins1, Bo Lönnerdal. 1. Department of Nutrition, the University of California, Davis, CA 95616, USA.
Abstract
BACKGROUND: Limited information exists on the biological role of a vitamin B-12-binding protein, haptocorrin, in human milk. The expression of haptocorrin by human mammary epithelial cells and its presence in human milk suggest a potential physiologic function in breastfed infants. OBJECTIVE: We investigated the extent to which haptocorrin could withstand proteolytic degradation and exert antimicrobial activity under in vitro conditions designed to simulate the gastrointestinal tract of breastfed infants. DESIGN: An in vitro model that simulates infant gastric and intestinal digestion was developed. The structural stability of porcine haptocorrin after exposure to digestive enzymes (pepsin and pancreatin) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, column chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The antimicrobial activity of haptocorrin was determined by incubating haptocorrin with enteropathogenic Escherichia coli O127 strain 2348/69 and monitoring bacterial growth. RESULTS: The structural analysis of haptocorrin exposed to enzymes did not show a decrease in molecular weight, which indicated that haptocorrin can survive proteolytic degradation. Both haptocorrin exposed to digestive enzymes and undigested haptocorrin inhibited the growth of enteropathogenic E. coli and did so to a similar extent. Thus, haptocorrin in vitro not only retains its structure after exposure to proteases but also exhibits antimicrobial activity. CONCLUSION: These results suggest that haptocorrin may exert a host-defense function against pathogens in the gastrointestinal tracts of breastfed infants.
BACKGROUND: Limited information exists on the biological role of a vitamin B-12-binding protein, haptocorrin, in human milk. The expression of haptocorrin by human mammary epithelial cells and its presence in human milk suggest a potential physiologic function in breastfed infants. OBJECTIVE: We investigated the extent to which haptocorrin could withstand proteolytic degradation and exert antimicrobial activity under in vitro conditions designed to simulate the gastrointestinal tract of breastfed infants. DESIGN: An in vitro model that simulates infant gastric and intestinal digestion was developed. The structural stability of porcine haptocorrin after exposure to digestive enzymes (pepsin and pancreatin) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, column chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The antimicrobial activity of haptocorrin was determined by incubating haptocorrin with enteropathogenic Escherichia coli O127 strain 2348/69 and monitoring bacterial growth. RESULTS: The structural analysis of haptocorrin exposed to enzymes did not show a decrease in molecular weight, which indicated that haptocorrin can survive proteolytic degradation. Both haptocorrin exposed to digestive enzymes and undigested haptocorrin inhibited the growth of enteropathogenic E. coli and did so to a similar extent. Thus, haptocorrin in vitro not only retains its structure after exposure to proteases but also exhibits antimicrobial activity. CONCLUSION: These results suggest that haptocorrin may exert a host-defense function against pathogens in the gastrointestinal tracts of breastfed infants.