Abstract
Human and bovine lactoferrin (hLf and bLf) are multifunctional iron-binding glycoprotein constitutively synthesized and secreted by glandular epithelial cells and by neutrophils following induction. HLf and bLf possess very high similarity of sequence. Therefore, most of the in vitro and in vivo studies are carried out with commercial bLf (cbLf), available in large quantities and recognized by Food and Drug Administration (FDA, USA) as a safe substance. Physico-chemical heterogeneity of different cbLf preparations influences their effectiveness. CbLf iron-saturation affects thermal stability and resistance to proteolysis. Moreover, other metal ions such as Al(III), Cu(II), Mg(II), Mn(II), Zn(II) are chelated by cbLf, even if at lower affinity than Fe(III). Ca(II) is also sequestered by the carboxylate groups of sialic acid present on glycan chains of cbLf thus provoking the release of LPS, contributing to bactericidal activity. Similarly to more than 50% of eukaryotic proteins, cbLf possesses five N-glycosylation sites, also contributing to the resistance to proteolysis and, putatively, to the protection of intestinal mucosa from pathogens. CbLfs possess several functions as anti-microbial, anti-biofilm, anti-adhesive, anti-invasive and anti-inflammatory activities. They are also relevant modulators of iron and inflammatory homeostasis. However, the efficacy of cbLfs in exerting several functions can be erratic mainly depending from integrity, degree of iron and other metal ions saturation, N-glycosylation sites and chains, desialylated forms, Ca(II) sequestration, presence of contaminants and finally the ability to enter inside nucleus.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abe H, Saito H, Miyakawa H, Tamura Y, Shimamura S, Nagao E, Tomita M (1991) Heat stability of bovine lactoferrin at acidic pH. J Dairy Sci 74:65–71
Ajello M, Greco R, Giansanti F, Massucci MT, Antonini G, Valenti P (2002) Anti-invasive activity of bovine lactoferrin towards group A streptococci. Biochem Cell Biol 80:119–124
Appelmelk BJ, An YQ, Geerts M, Thijs BG, de Boer HA, MacLaren DM, de Graaff J, Nuijens JH (1994) Lactoferrin is a lipid A-binding protein. Infect Immun 62:2628–2632
Armstead AL, Li B (2011) Nanomedicine as an emerging approach against intracellular pathogens. Int J Nanomed 6:3281–3293. https://doi.org/10.2147/IJN.S27285
Ashida K, Sasaki H, Suzuki YA, Lönnerdal B (2004) Cellular internalization of lactoferrin in intestinal epithelial cells. Biometals 17:311–315
Baker E (1994) Structure and reactivity of transferrins. Adv Inorg Chem 41:389–463
Baker EN, Baker HM (2005) Molecular structure, binding properties and dynamics of lactoferrin. Cell Mol Life Sci 62:2531–2539
Baker EN, Baker HM (2009) A structural framework for understanding the multifunctional character of lactoferrin. Biochimie 91:3–10. https://doi.org/10.1016/j.biochi.2008.05.006
Barboza M, Pinzon J, Wickramasinghe S, Froehlich JW, Moeller I, Smilowitz JT, Ruhaak LR, Huang J, Lönnerdal B, German JB (2012) Glycosylation of human milk lactoferrin exhibits dynamic changes during early lactation enhancing its role in pathogenic bacteria-host interactions. Mol Cell Proteom 11(M111):015248. https://doi.org/10.1074/mcp.M111.015248
Bellamy W, Takase M, Wakabayashi H, Kavase K, Tomita M (1992) Antibacterial spectrum of lactoferricin B, a potent bactericide peptide derived from the N-terminal region of bovine lactoferrin. J Appl Bacteriol 73:472–479
Berlutti F, Morea C, Battistoni A, Sarli S, Cipriani P, Superti F, Ammendolia MG, Valenti P (2005) Iron availability influences aggregation, biofilm, adhesion and invasion of Pseudomonas aeruginosa and Burkholderia cenocepacia. Int J Immunopathol Pharmacol l18:661–67. https://doi.org/10.1177/039463200501800407
Berlutti F, Schippa S, Morea C, Sarli S, Perfetto B, Donnarumma G, Valenti P (2006) Lactoferrin downregulates pro-inflammatory cytokines up-expressed in intestinal epithelial cells infected with invasive or noninvasive Escherichia coli strains. Biochem Cell Biol 84:351–357. https://doi.org/10.1139/o06-039
Berlutti F, Superti F, Nicoletti M, Morea C, Frioni A, Ammendolia MG, Battistoni A, Valenti P (2008) Bovine lactoferrin inhibits the efficiency of invasion of respiratory A549 cells of different iron-regulated morphological forms of Pseudomonas aeruginosa and Burkholderia cenocepacia. Int J Immunopathol Pharmacol 21:51–59. https://doi.org/10.1177/039463200802100107
Berlutti F, Catizone A, Ricci G, Frioni A, Natalizi T, Valenti P, Polimeni A (2010) Streptococcus mutans and Streptococcus sobrinus are able to adhere and invade human gingival fibroblast cell line. Int J Immunopathol Pharmacol 23:1253–1260. https://doi.org/10.1177/039463201002300430
Berlutti F, Pantanella F, Natalizi T, Frioni A, Paesano R, Polimeni A, Valenti P (2011) Antiviral properties of lactoferrin—a natural immunity molecule. Molecules 16:6992–7018. https://doi.org/10.3390/molecules16086992
Brandenburg K, Jurgens G, Muller M, Fukuoka S, Koch MHJ (2001) Biophysical characterization of lipopolysaccharide and lipid A inactivation by lactoferrin. Biol Chem 382:1215–1225
Brines R, Brock J (1983) The effect of trypsin and chymotrypsin on the in vitro antimicrobial and iron-binding properties of lactoferrin in human milk and bovine colostrum: unusual resistance of human apolactoferrin to proteolytic digestion. Biochim Biophys 759:229–235. https://doi.org/10.1016/0304-4165(83)90317-3
Cario E, Rosenberg IM, Brandwein SL, Beck PL, Reinecker HC, Podolsky DK (2000) Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing Toll-like receptors. J Immunol 164:966–972
Chanda W, Joseph TP, Wang W, Padhiar AA, Zhong M (2017) The potential management of oral candidiasis using anti-biofilm therapies. Med Hypotheses 106:15–18. https://doi.org/10.1016/j.mehy.2017.06.029
Cutone A, Frioni A, Berlutti F, Valenti P, Musci G, Bonaccorsi di Patti MC (2014) Lactoferrin prevents LPS-induced decrease of the iron exporter ferroportin in human monocytes/macrophages. Biometals 27:807–813. https://doi.org/10.1007/s10534-014-9742-7
Cutone A, Rosa L, Lepanto MS, Scotti MJ, Berlutti F, Bonaccorsi di Patti MC, Musci G, Valenti P (2017) Lactoferrin efficiently counteracts the inflammation-induced changes of the iron homeostasis system in macrophages. Front Immunol 8:705. https://doi.org/10.3389/fimmu.2017.00705
Frioni A, Conte MP, Cutone A, Longhi C, Musci G, di Patti MC, Natalizi T, Marazzato M, Lepanto MS, Puddu P, Paesano R, Valenti P, Berlutti F (2014) Lactoferrin differently modulates the inflammatory response in epithelial models mimicking human inflammatory and infectious diseases. Biometals 27:843–856. https://doi.org/10.1007/s10534-014-9740-9
Goldoni P, Sinibaldi L, Valenti P, Orsi N (2000) Metal complexes of lactoferrin and their effect on the intracellular multiplication of Legionella pneumophila. Biometals 13:15–22
Groves ML (1960) The isolation of a red protein from milk. J Am Chem Soc 82:3345–3350. https://doi.org/10.1021/ja01498a700
Hayworth JL, Kasper KJ, Leon-Ponte M, Herfst CA, Yue D, Brintnell WC, Mazzuca DM, Heinrichs DE, Cairns E, Madrenas J, Hoskin DW, McCormick JK, Haeryfar SM (2009) Attenuation of massive cytokine response to the staphylococcal enterotoxin B superantigen by the innate immunomodulatory protein lactoferrin. Clin Exp Immunol 157:60–70. https://doi.org/10.1111/j.1365-2249.2009.03963.x
Jabeen T, Sharma S, Singh N, Bhushan A, Singh TP (2005) Structure of the zinc-saturated C-terminal lobe of bovine lactoferrin at 2.0 A resolution. Acta Crystallogr D Biol Crystallogr 61:1107–1115. https://doi.org/10.1107/S0907444905016069
Ji ZS, Mahley RW (1994) Lactoferrin binding to heparan sulfate proteoglycans and the LDL receptor-related protein. Further evidence supporting the importance of direct binding of remnant lipoproteins to HSPG. Arterioscler Thromb Vasc Biol 14:2025–2031
Johansson B (1960) Isolation of an iron-containing red protein from human milk. Acta Chem Scand 14:510–512
Karav S, German JB, Rouquié C, Le Parc A, Barile D (2017) Studying lactoferrin N-glycosylation. Int J Mol Sci 18(4). https://doi.org/10.3390/ijms18040870
Kautto L, Nguyen-Khuong T, Everest-Dass A, Leong A, Zhao Z, Willcox MD, Packer NH, Peterson R (2016) Glycan involvement in the adhesion of Pseudomonas aeruginosa to tears. Exp Eye Res 145:278–288. https://doi.org/10.1016/j.exer.2016.01.013
Kim CW, Lee TH, Park KH, Choi SY, Kim J (2012) Human lactoferrin suppresses TNF-a-induced intercellular adhesion molecule-1 expression via competition with NF-kB in endothelial cells. FEBS Lett 586:229–234. https://doi.org/10.1016/j.febslet.2011.12.011
Latorre D, Berlutti F, Valenti P, Gessani S, Puddu P (2012) LF immunomodulatory strategies: mastering bacterial endotoxin. Biochem Cell Biol 90:269–278. https://doi.org/10.1139/o11-059
Legrand D (2012) Lactoferrin, a key molecule in immune and inflammatory processes. Biochem Cell Biol 90:252–268. https://doi.org/10.1139/o11-056
Legrand D, Elass E, Carpentier M, Mazurier J (2005) Lactoferrin: a modulator of immune and inflammatory responses. Cell Mol Life Sci 62:2549–2559. https://doi.org/10.1007/s00018-005-5370-2
Lonnerdal B (2009) Nutritional roles of lactoferrin. Curr Opin Clin Nutr Metab Care 12:293–297. https://doi.org/10.1097/MCO.0b013e328328d13e
Masson PL, Heremans JF (1971) Lactoferrin in milk from different species. Comp Biochem Physiol 39:119–129
Masson PL, Heremans JF, Dive C (1966) An iron-binding protein common to many external secretions. Clin Chim Acta 14:735–739
Masson PL, Heremans JF, Schonne E (1969) Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J Exp Med 130:643–658
Montreuil J, Tonnelat J, Mullet S (1960) Preparation and properties of lactosiderophilin (lactotransferrin) of human milk. Biochim Biophys Acta 45:413–421
Moore SA, Anderson BF, Groom CR, Haridas M, Baker EN (1997) Three-dimensional structure of diferric bovine lactoferrin at 2.8 A resolution. J Mol Biol 274:222–236. https://doi.org/10.1006/jmbi.1997.1386
Paesano R, Torcia F, Berlutti F, Pacifici E, Ebano V, Moscarini M, Valenti P (2006) Oral administration of lactoferrin increases hemoglobin and total serum iron in pregnant women. Biochem Cell Biol 84:377–380. https://doi.org/10.1139/o06-040
Paesano R, Pietropaoli M, Gessani S, Valenti P (2009) The influence of lactoferrin, orally administered, on systemic iron homeostasis in pregnant women suffering of iron deficiency and iron deficiency anaemia. Biochimie 91:44–51. https://doi.org/10.1016/j.biochi.2008.06.004
Paesano R, Berlutti F, Pietropaoli M, Pantanella F, Pacifici E, Goolsbee W, Valenti P (2010) Lactoferrin efficacy versus ferrous sulfate in curing iron deficiency and iron deficiency anemia in pregnant women. Biometals 23:411–417. https://doi.org/10.1177/039463201002300220
Paesano R, Natalizi T, Berlutti F, Valenti P (2012) Body iron delocalization: the serious drawback in iron disorders in both developing and developed countries. Pathog Glob Health 106:200–216. https://doi.org/10.1179/2047773212Y.0000000043
Paesano R, Pacifici E, Benedetti S, Berlutti F, Frioni A, Polimeni A, Valenti P (2014) Safety and efficacy of lactoferrin versus ferrous sulphate in curing iron deficiency and iron deficiency anaemia in hereditary thrombophilia pregnant women: an interventional study. Biometals 27:999–1006. https://doi.org/10.1007/s10534-014-9723-x
Puddu P, Carollo MG, Belardelli F, Valenti P, Gessani S (2007) Role of endogenous interferon and LPS in the immunomodulatory effects of bovine lactoferrin in murine peritoneal macrophages. J Leukoc Biol 82:347–353. https://doi.org/10.1189/jlb.1106688
Puddu P, Valenti P, Gessani S (2009) Immunomodulatory effects of lactoferrin on antigen presenting cells. Biochimie 91:11–18. https://doi.org/10.1016/j.biochi.2008.05.005
Puddu P, Latorre D, CarolloM Catizone A, Ricci G, Valenti P, Gessani S (2011) Bovine lactoferrin counteracts Toll-like receptor mediated activation signals in antigen presenting cells. PLoS ONE 6:e22504. https://doi.org/10.1371/journal.pone.0022504
Rosa L, Cutone A, Lepanto MS, Paesano R, Valenti P (2017) Lactoferrin: a natural glycoprotein involved in iron and inflammatory homeostasis. Int J Mol Sci. https://doi.org/10.3390/ijms18091985
Rossi P, Giansanti F, Boffi A, Ajello M, Valenti P, Chiancone E, Antonini G (2002) Ca2+ binding to bovine lactoferrin enhances protein stability and influences the release of bacterial lipopolysaccharide. Biochem Cell Biol 80:41–48
Sessa R, Di Pietro M, Filardo S, Bressan A, Rosa L, Cutone A, Frioni A, Berlutti F, Paesano R, Valenti P (2017) Effect of bovine lactoferrin on Chlamydia trachomatis infection and inflammation. Biochem Cell Biol 95:34–40. https://doi.org/10.1139/bcb-2016-0049
Shin K, Wakabayashi H, Yamauchi K, Yaeshima T, Iwatsuki K (2008) Recombinant human intelectin binds bovine lactoferrin and its peptides. Biol Pharm Bull 31:1605–1608
Siciliano R, Rega B, Marchetti M, Seganti L, Antonini G, Valenti P (1999) Bovine lactoferrin peptidic fragments involved in inhibition of herpes simplex virus type 1 infection. Biochem Biophys Res Commun 264:19–23. https://doi.org/10.1006/bbrc.1999.1318
Sorensen M, Sorensen S (1940) The proteins in whey. Compte rendu des Travaux du Laboratoire de Carlsberg Ser. Chim 23:55–99
Stallmann S, Hegemann JH (2015) The Chlamydia trachomatis Ctad1 invasin exploits the human integrin 1 receptor for host cell entry. Cell Microbiol 18:761–775. https://doi.org/10.1111/cmi.12549
Steijns JM, van Hooijdonk AC (2000) Occurrence, structure, biochemical properties and technological characteristics of lactoferrin. Br J Nutr 1:11–17
Superti F, Ammendolia MG, Valenti P, Seganti L (1997) Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29. Med Microbiol Immunol 186:83–91
Superti F, Siciliano R, Rega B, Giansanti F, Valenti P, Antonini G (2001) Involvement of bovine lactoferrin metal saturation, sialic acid and protein fragments in the inhibition of rotavirus infection. Biochim Biophys Acta 1528:107–115
Suzuki YA, Wong H, Ashida KY, Schryvers AB, Lönnerdal B (2008) The N1 domain of human lactoferrin is required for internalization by Caco-2 cells and targeting to the nucleus. Biochemistry 47:10915–10920. https://doi.org/10.1021/bi8012164
Tomita M, Bellamy W, Takase M, Yamauchi K, Wakabayashi K, Kavase K (1991) Potent antibacterial peptides generated by pepsin of bovine lactoferrin. J Dairy Sci 74:4137–4142. https://doi.org/10.3168/jds.S0022-0302(91)78608-6
Valenti P, Antonini G (2005) Lactoferrin: an important host defence against microbial and viral attack. Cell Mol Life Sci 62:2576–2587. https://doi.org/10.1007/s00018-005-5372-0
Valenti P, Catizone A, Pantanella F, Frioni A, Natalizi T, Tendini M, Berlutti F (2011) Lactoferrin decreases inflammatory response by cystic fibrosis bronchial cells invaded with Burkholderia cenocepacia iron-modulated biofilm. Int J Immunopathol Pharmacol 24:1057–1068. https://doi.org/10.1177/039463201102400423
Valenti P, Frioni A, Rossi A, Ranucci S, De Fino I, Cutone A, Rosa L, Bragonzi A, Berlutti F (2017) Aerosolized bovine lactoferrin reduces neutrophils and pro-inflammatory cytokines in mouse models of Pseudomonas aeruginosa lung infections. Biochem Cell Biol 95:41–47. https://doi.org/10.1139/bcb-2016-0050
VanVeen HA, Geerts ME, van Berkel PH, Nuijens JH (2004) The role of n-linked glycosylation in the protection of human and bovine lactoferrin against tryptic proteolysis. Eur J Biochem 271:678–684. https://doi.org/10.1111/j.1432-1033.2003.03965.x
Visca P, Berlutti F, Vittorioso P, Dalmastri C, Thaller MC, Valenti P (1989) Growth and adsorption of Streptococcus mutans 6715–13 to hydroxyapatite in the presence of lactoferrin. Med Microbiol Immunol 178:69–79
Vora P, Youdim A, Thomas LS, Fukata M, Tesfay SY, Lukasek K, Michelsen KS, Wada A, Hirayama T, Arditi M, Abreu MT (2004) Beta-defensin-2 expression is regulated by TLR signaling in intestinal epithelial cells. J Immunol 173:5398–5405
Ward PP, Paz E, Conneely OM (2005) Multifunctional roles of lactoferrin: a critical overview. Cell Mol Life Sci 62:2540–2548. https://doi.org/10.1007/s00018-005-5369-8
Wormald MR, Petrescu AJ, Pao Y-L, Glithero A, Elliott T, Dwek RA (2002) Conformational studies of oligosaccharides and glycopeptides: complementarity of NMR, X-ray crystallography, and molecular modelling. Chem Rev 102:371–386
Wu HF, Monroe DM, Church FC (1995) Characterization of the glycosaminoglycan-binding region of Lactoferrin. Arch Biochem Biophys 317:85–92. https://doi.org/10.1006/abbi.1995.1139
Acknowledgements
This work was granted by University of Rome La Sapienza Funds to PV.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Rights and permissions
About this article
Cite this article
Rosa, L., Cutone, A., Lepanto, M.S. et al. Physico-chemical properties influence the functions and efficacy of commercial bovine lactoferrins. Biometals 31, 301–312 (2018). https://doi.org/10.1007/s10534-018-0092-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10534-018-0092-8