Glycation Ligand Binding Motif in Lactoferrin

Implications in Diabetic Infection
  • Yong Ming Li
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 443)


Lactoferrin and lysozyme are two important, naturally occurring antibacterial proteins found in saliva, nasal secretions, milk, mucus, serum and in the lysosomes of neutrophils and macrophages. Both proteins bind specifically to glucose-modified proteins bearing advanced glycation endproducts (AGEs). Exposure to AGE-modified proteins blocks the bacterial agglutination and bacterial killing activities of lactoferrin and also inhibits the bactericidal and enzymatic activity of lysozyme. Peptide mapping by AGE ligand blot revealed two AGE-binding domains in lactoferrin, and a single AGE-binding domain in lysozyme. None of these AGE-binding domains displayed any significant homology in their primary sequences; however, a common 17–18 amino acid cysteine loop motif (CX15–16C) was identified among them, which we named an ABCD motif (AGE-Binding Cysteine-bounded Domain). Similar domains are also present in other antimicrobial proteins such as defesins. Hydrophilicity analysis indicated that each of these ABCD loops is markedly hydrophilic. Synthetic peptides, corresponding to these motifs in lactoferrin and lysozyme, exhibited AGE-binding activity. Since diabetes is associated with abnormally high levels of tissue and serum AGEs, the elevated AGEs may inhibit endogenous antibacterial proteins by binding to the conserved ABCD motif, thereby increasing susceptibility to bacterial infections in diabetic individuals. These results may provide a basis for the development of new approaches to prevent diabetic infections.


Minimal Inhibitory Concentration Defense Protein Bovine Lactoferrin Cidal Domain Bacterial Agglutination 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Raphael GD, Jeney EV, Baraniuk JN, Kim I, Meredith SD, Kaliner MA. Pathophysiology of rhinitis. lactoferrin and lysozyme in nasal secretions. J Clin Invest 1989; 84 (5): 1528–35.PubMedCrossRefGoogle Scholar
  2. 2.
    Taylor DC, Cripps AW, Clancy RL. A possible role for lysozyme in determining acute exacerbation in chronic bronchitis. Clin Exp Immunol 1995; 102 (2): 406–16.PubMedCrossRefGoogle Scholar
  3. 3.
    Adeyemi E0, D’Anastasio C, Impallomeni MG, Hodgson HJ. Plasma lactoferrin as a marker of infection in elderly individuals. Aging (Milano) 1992; 4 (2): 135–7.Google Scholar
  4. 4.
    Bellamy W, Takase M, Wakabayashi H, Kawase K, Tornita M. Antibacterial spectrum of lactoferricin b, a potent bactericidal peptide derived from the N-terminal region of bovine lactoferrin. J Appl Bacteriol 1992; 73 (6): 472–9.PubMedCrossRefGoogle Scholar
  5. 5.
    Longhi C, Conte MP, Seganti L, Polidoro M, Alfsen A, Valenti P. Influence of lactoferrin on the entry process of Escherichia coli hbl0l (pri203) in HELA cells. Med Microbiol Immunol (Berl) 1993; 182 (1): 25–35.CrossRefGoogle Scholar
  6. 6.
    Bellamy W, Wakabayashi H, Takase M, Kawase K, Shimamura S, Tornita M. Killing of Candida albicans by lactoferricin b, a potent antimicrobial peptide derived from the N-terminal region of bovine lactoferrin. Med Microbiol Immunol (Berl) 1993; 182 (2): 97–105.CrossRefGoogle Scholar
  7. 7.
    Byrd TF, Horwitz MA. Lactoferrin inhibits or promotes Legionella pneumophila intracellular multiplication in nonactivated and interferon gamma-activated human monocytes depending upon its degree of iron saturation. Iron-lactoferrin and nonphysiologic iron chelates reverse monocyte activation against Legionella pneumophila. J Clin Invest 1991; 88 (4): 1103–12.PubMedCrossRefGoogle Scholar
  8. 8.
    Miehlke S, Reddy R, Osato MS, Ward PP, Conneely OM, Graham DY. Direct activity of recombinant human lactoferrin against Helicohacter pylori. J Clin Microbiol 1996; 34 (10): 2593–4.PubMedGoogle Scholar
  9. 9.
    Marchetti M, Longhi C, Conte MP, Pisani S, Valenti P, Seganti L. Lactoferrin inhibits Herpes simplex virus type 1 adsorption to Vero cells. Antiviral Res 1996; 29 (2–3): 221–31.PubMedCrossRefGoogle Scholar
  10. 10.
    Shimizu K, Matsuzawa I, Okada K, Tazume S, Dosako S, Kawasaki Y, Hashimoto K, Koga Y. Lactoferrin-mediated protection of the host from murine cytomegalovirus infection by a T-cell-dependent augmentation of natural killer cell activity. Arch Virol 1996; 141 (10): 1875–89.PubMedCrossRefGoogle Scholar
  11. I I. Tanaka T, Ornata Y, Saito A, Shimazaki K, Igarashi I, Suzuki N. Growth inhibitory effects of bovine lactoferrin to Toxoplasma gondii parasites in murine somatic cells. J Vet Med Sci 1996; 58 (1): 61–5.PubMedCrossRefGoogle Scholar
  12. 12.
    Miyazawa K, Mantel C, Lu L, Morrison DC, Broxmeyer HE. Lactoferrin-lipopolysaccharide interactions. Effect on lactoferrin binding to monocyte/macrophage-differentiated HL-60 cells. J Immunol 1991; 146 (2): 723–9.PubMedGoogle Scholar
  13. 13.
    Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K, Tomita M. Identification of the bactericidal domain of lactoferrin. Biochim Biophys Acta 1992; 1121 (I-2): 130–6.PubMedCrossRefGoogle Scholar
  14. 14.
    Cerami A, Vlassara H, Brownlee M. Glucose and aging. Sci Am 1987; 256 (5): 90–6.PubMedCrossRefGoogle Scholar
  15. 15.
    Bucala R, Vlassara H. Advanced glycosylation endproducts in diabetic renal disease: clinical measurement, pathophysiological significance, and prospects for pharmacological inhibition. Blood Purif 1995: 13 (3–4): 160–70.PubMedCrossRefGoogle Scholar
  16. 16.
    Li YM, Steffes M, Donnelly T, Liu C, Fuh H, Basgen J, Bucala R, Vlassara H. Prevention of cardiovascular and renal pathology of aging by the advanced glycation inhibitor aminoguanidine. Proc Natl Acad Sci U S A 1996; 93 (9): 3902–7.PubMedCrossRefGoogle Scholar
  17. 17.
    Vlassara H, Bucala R, Striker L. Pathogenic effects of advanced glycosylation: biochemical, biologic, and clinical implications for diabetes and aging. Lab Invest 1994; 70 (2): 138–51.PubMedGoogle Scholar
  18. 18.
    Li YM, Baviello G, Vlassara H, Mitsuhashi T. Glycation products in aged thioglycollate medium enhance the elicitation of peritoneal macrophages. J Immunol Meth 1997; 201: 183–8.CrossRefGoogle Scholar
  19. 19.
    Li YM, Dickson DW. Enhanced binding of advanced glycation endproducts (AGE) by the ApoE4 isoform links the mechanism of plaque deposition in Alzheimer’s disease. Neuroscience Lett 1997; 226: 155–158.CrossRefGoogle Scholar
  20. 20.
    Vlassara H, Brownlee M, Cerami A. Novel macrophage receptor for glucose-modified proteins is distinct from previously described scavenger receptors. J Exp Med 1986; 164 (4): 1301–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Yang Z, Makita Z, Horii Y, Brunelle S, Cerami A, Sehajpal P, Suthanthiran M, Vlassara H. Two novel rat liver membrane proteins that bind advanced glycosylation endproducts: relationship to macrophage receptor for glucose-modified proteins. J Exp Med 1991; 174 (3): 515–24.PubMedCrossRefGoogle Scholar
  22. 22.
    Li YM, Mitsuhashi T, Wojciechowicz D, Shimizu N, Li J, Stitt A, He C, Banerjee D, Vlassara H. Molecular identity and cellular distribution of advanced glycation endproduct receptors: relationship of p60 to ost48 and p90 to 80k-h membrane proteins. Proc Natl Acad Sci U S A 1996; 93 (20): 11047–52.PubMedCrossRefGoogle Scholar
  23. 23.
    Neeper M, Schmidt AM, Brett J, Yan SD, Wang F. Pan YC, Elliston K, Stem D, Shaw A. Cloning and expression of a cell surface receptor for advanced glycosylation end products of proteins. J Biol Chem 1992; 267 (21): 14998–5004.PubMedGoogle Scholar
  24. 24.
    Schmidt AM, Vianna M, Gerlach M, Brett J, Ryan J. Kao J, Esposito C, Hegarty H, Hurley W, Clauss M, et al. Isolation and characterization of two binding proteins for advanced glycosylation end products from bovine lung which are present on the endothelial cell surface. J Biol Chem 1992; 267 (21): 14987–97.PubMedGoogle Scholar
  25. 25.
    Li YM, Tan AX, Vlassara H. Antibacterial activity of lysozyme and lactoferrin is inhibited by binding of advanced glycation-modified proteins to a conserved motif. Nat Med 1995; 1 (10): 1057–61.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Yong Ming Li
    • 1
  1. 1.The Picower Institute for Medical ResearchManhassetUSA

Personalised recommendations