Influences of Animal Mucins on Peroxidase Activity in Solution and on the Surface of Hydroxyapatite

동물성 Mucin이 용액상태와 Hydroxyapatite표면에서 Peroxidase 활성에 미치는 영향에 관한 연구

  • Lee, Sang-Goo (Dept. of Oral Medicine & Oral Diagnosis, School of Dentistry & Dental Research Institute, Seoul National University) ;
  • Jeon, Eun-Hyoung (Dept. of Oral Medicine & Oral Diagnosis, School of Dentistry & Dental Research Institute, Seoul National University) ;
  • Kho, Hong-Seop (Dept. of Oral Medicine & Oral Diagnosis, School of Dentistry & Dental Research Institute, Seoul National University)
  • 이상구 (서울대학교 치과대학 구강내과진단학 교실, 치학연구소) ;
  • 전은형 (서울대학교 치과대학 구강내과진단학 교실, 치학연구소) ;
  • 고홍섭 (서울대학교 치과대학 구강내과진단학 교실, 치학연구소)
  • Published : 2008.09.30

Abstract

Animal mucins have structural characteristics similar to human salivary mucins. Animal mucins have been regarded as suitable substances for saliva substitutes. Since animal mucin molecules in saliva substitutes and host-derived antimicrobial salivary molecules exist simultaneously in whole saliva and the pellicles of patients with dry mouth, interactions may occur between these molecules. The purpose of this study was to investigate the influence of animal mucins on peroxidase activity in solution and on the surface of hydroxyapatite(HA) surfaces. The effects of animal mucins on peroxidase activity were examined by incubating porcine gastric mucin(PGM) or bovine submaxillary mucin (BSM) with either bovine lactoperoxidase(bLPO) or saliva samples. For solid-phase assays, immobilized animal mucins or peroxidase on three different HA surfaces(HA beads, HA disc, and bovine tooth) were used. Peroxidase activity was determined with an NbsSCN assay. The obtained results were as follows: 1. PGM enhanced the enzymatic activity of bLPO in solution phase. PGM did not affect the enzymatic activity of peroxidase in saliva sample(POS). 2. BSM did not affect the enzymatic activities of both bLPO and POS in solution phase. 3. HA-adsorbed PGM increased subsequent bLPO adsorption in all three HA phases. The activity of POS was increased on both the HA beads and bovine tooth. 4. The peroxidase activities on the HA beads and disc were increased when the HA surfaces were exposed to a mixture of bLPO and PGM. 5. The binding affinity of bLPO to PGM was greater than that of bLPO to BSM. Collectively, our results suggest that animal mucins affects the enzymatic activity of peroxidase on the HA surfaces as well as in solution. Saliva substitutes containing animal mucins may affect the function of antimicrobial components in natural saliva and saliva substitutes.

Keywords

Porcine gastric mucin;Bovine submaxillary mucin;Peroxidase;Saliva

References

  1. Cohen RE, Levine MJ. Salivary glycoproteins. In: Tenovuo JO, ed. Human Saliva: Clinical Chemistry and Microbiology. Vol I. Boca Raton, 1989, CRC Press Inc., pp. 101-30
  2. Mellema J, Holterman HJ, Waterman HA, Blom C, 'S-Gravenmade EJ. Rheological aspects of mucin-containing solutions and saliva substitutes. Biorheology 1992;29:231-249 https://doi.org/10.3233/BIR-1992-292-304
  3. Vissink A, 'S-Gravenmade EJ, Panders AK, Vermey A, Petersen JK, Visch LL, Schaub RMH. A clinical comparison between commercially available mucinand CMC-containing saliva substitutes. Int J Oral Surg 1983;12:232-238 https://doi.org/10.1016/S0300-9785(83)80048-9
  4. Visch LL, 'S-Gravenmade EJ, Schaub RMH, Van Putten WLJ, Vissink A. A double-blind crossover trial CMC- and mucin-containing saliva substitutes. Int J Oral Surg 1986;15:395-400 https://doi.org/10.1016/S0300-9785(86)80027-8
  5. Duxbury AJ, Thakker NS, Wastell DG. A double-blind cross-over trial of a mucin-containing artificial saliva. Br Dent J 1989;166:115-120 https://doi.org/10.1038/sj.bdj.4806731
  6. Turner BS, Bhaskar KR, Hadzopoulou-Cladaras M, Specian RD, LaMont JT. Isolation and characterization of cDNA clones encoding pig gastric mucin. Biochem J 1995;308:89-96 https://doi.org/10.1042/bj3080089
  7. Roger V, Tenovuo J, Lenander Lumikari M, Sderling E, Vilja P. Lysozyme and lactoperoxidase inhibit the adherence of Streptococcus mutans NCTC 10449 (serotype c) to saliva treated hydroxyapatite in vitro. Caries Res 1994;28:421-428 https://doi.org/10.1159/000262015
  8. Schilling KM, Bowen WH. The activity of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite. J Dent Res 1988;67:2-8 https://doi.org/10.1177/00220345880670010201
  9. Biesbrock AR, Reddy MS, Levine MJ. Interaction of a salivarymucin-secretory immunoglobulin A complex with mucosal pathogens. Infect Immun 1991;59:3492-3497
  10. Loomis RE, Prakobphol A, Levine MJ, Reddy MS, Jones PC. Biochemical and biophysical comparison of two mucins from human submandibular-sublingual saliva. Arch Biochem Biophys 1987;258:452-464 https://doi.org/10.1016/0003-9861(87)90366-3
  11. Hannig M. Transmission electron microscopic study of in vivo pellicle formation of dental restorative materials. Eur J Oral Sci 1997;105:422-433 https://doi.org/10.1111/j.1600-0722.1997.tb02139.x
  12. Hannig C, Hoch J, Becker K, Hannig M, Attin T. Lysozyme activity in the initially formed in situ pellicle. Arch Oral Biol 2005;50:821-828 https://doi.org/10.1016/j.archoralbio.2005.01.006
  13. Vissink A, Schaub RMH, Van Rijn LJ, 'S-Gravenmade EJ, Panders AK, Vermey A. The efficacy of mucin-containing artificial saliva in alleviating symptoms of xerostomia. Gerodontology 1987;6:95-101 https://doi.org/10.1111/j.1741-2358.1987.tb00283.x
  14. Yao Y, Grogan J, Zehnder M, Lendenmann U, Nam B, Wu Z, Costello CE, Oppenheim FG. Compositional analysis of human acquired pellicle by mass spectrometry. Arch Oral Biol 2001;46:293-303 https://doi.org/10.1016/S0003-9969(00)00134-5
  15. Soares RV, Siqueira CC, Bruno LS, Oppenheim FG, Offner GD, Troxler RF. MG2 and lactoferrin form a heterotypic complex in salivary secretions. J Dent Res 2003;82:471-475 https://doi.org/10.1177/154405910308200613
  16. Pigman W, Moschera J, Weis M, Tettamanti G. The occurrence of repetitive glycopeptide sequences in bovine submaxillary glycoprotein. Eur J Biochem 1973;32:148-154 https://doi.org/10.1111/j.1432-1033.1973.tb02591.x
  17. Park WK, Chung JW, Kim YK, Chung SC, Kho HS. Influences of animal mucins on lysozyme activity in solution and on hydroxyapatite surfaces. Arch Oral Biol 2006;51:861-869 https://doi.org/10.1016/j.archoralbio.2006.04.002
  18. Hannig M. Ultrastructural investigation of pellicle morphogenesis at two different intraoral sites during 24-h period. Clin Oral Investig 1999;3:88-95 https://doi.org/10.1007/s007840050084
  19. Tabak LA, Levine MJ, Jain NK, Bryan AR, Cohen RE, Monte LD, Zawacki S, Nancollas GH, Slomiany A, Slomiany BL. Adsorption of human salivary mucins to hydroxyapatite. Arch Oral Biol 1985;30:423-427 https://doi.org/10.1016/0003-9969(85)90070-6
  20. Al-Hashimi I, Levine MJ. Characterization of in vivo salivary-derived enamel pellicle. Arch Oral Biol 1989;34:289-295 https://doi.org/10.1016/0003-9969(89)90070-8
  21. Stayton PS, Drobny GP, Shaw WJ, Long JR, Gilbert M. Molecular recognition at the protein-hydroxyapatite interface. Crit Rev Oral Biol Med 2003;14:370-376 https://doi.org/10.1177/154411130301400507
  22. Busscher HJ, Uyen HM, Stokroos I, Jongebloed WL. A transmission electron microscopy study of the adsorption patterns of early developing artificial pellicles on human enamel. Arch Oral Biol 1989;34:803-810 https://doi.org/10.1016/0003-9969(89)90031-9
  23. Snary D, Allen A, Pain RH. Structural studies on gastric mucoproteins; lowering of molecular weight after reduction with 2-mercaptoethanol. Biochem Biophys Res Commun 1970;40:844-851 https://doi.org/10.1016/0006-291X(70)90980-0
  24. Pruitt KM, Caldwell RC, Jamieson AD, Taylor RE. The interaction of salivary proteins with tooth surface. J Dent Res 1969;48:818-823 https://doi.org/10.1177/00220345690480053501
  25. Rlla G, Ciardi JE, Bowen WH. Identification of IgA, IgG, lysozyme, albumin, ${\alpha}$-amylase and glucosyltransferase in the protein layer adsorbed to hydroxyapatite from whole saliva. Scand J Dent Res 1983;91:186-190
  26. Dodds MWJ, Johnson DA, Yeh C. Health benefits of saliva: a review. J Dent 2005;33:223-233 https://doi.org/10.1016/j.jdent.2004.10.009
  27. Hannig C, Hannig M, Attin T. Enzymes in the acquired enamel pellicle. Eur J Oral Sci 2005;113:2-13 https://doi.org/10.1111/j.1600-0722.2004.00180.x
  28. van der Mei HC, White DJ, Kamminga-Rasker HJ, Knight J, Baig AA, Smit J, Busscher HJ. Influence of dentifrices and dietary components in saliva on wettability of pellicle-coated enamel on vitro and in vivo. Eur J Oral Sci 2002;110:434-438 https://doi.org/10.1034/j.1600-0722.2002.21341.x
  29. Carlen A, Brjesson AC, Nikdel K, Olsson J. Composition of pellicles formed in vivo on tooth surfaces in different parts of the dentition, and in vitro on hydroxyapatite. Caries Res 1998;32:447-455 https://doi.org/10.1159/000016486
  30. Park MS, Chung JW, Kim YK, Chung SC, Kho HS. Viscosity and wettability of animal mucin solutions and human saliva. Oral Dis 2007;13:181-186 https://doi.org/10.1111/j.1601-0825.2006.01263.x
  31. Schenkels LCPM, Gururaja TL, Levine MJ. Salivary mucins: Their role in oral mucosal barrier function and drug delivery. In Rathbone MJ (ed). Oral mucosal drug delivery. New York, 1996, Marcel Dekker, pp. 191-220
  32. Tenovuo J, Kurkijrvi K. Immobilized lactoperoxidase as a biologically active and stable form of an antimicrobial enzyme. Arch Oral Biol 1981;26:309-314 https://doi.org/10.1016/0003-9969(81)90052-2
  33. O'Brien PJ. Peroxidases. Chem Biol Interact 2000;129:113-139 https://doi.org/10.1016/S0009-2797(00)00201-5
  34. Vacca-Smith AM, Venkitaraman AR, Schilling KM, Bowen WH. Characterization of glucosyltransferase of human saliva adsorbed on to hydroxyapatite surfaces. Caries Res 1996;30:354-360 https://doi.org/10.1159/000262342
  35. Prakobphol A, Levine MJ, Tabak LA, Reddy MA. Purification of a low-molecular weight mucin type glycoprotein from human submandibular-sublingual saliva. Carbohydr Res 1982;108:111-122 https://doi.org/10.1016/S0008-6215(00)81896-0
  36. Slomiany BL, Murty VLN, Piotrowski J, Slomiany A. Salivary mucins in oral mucosal defense. Gen Pharmac 1996;27:761-771 https://doi.org/10.1016/0306-3623(95)02050-0
  37. Vacca-Smith AM, Venkitaraman AR, Quivey RG, Bowen WH. Interactions of streptococcal glucosyltransferases with ${\alpha}$-amylase and starch on the surface of saliva-coated hydroxyapatite. Arch Oral Biol 1996;41:291-298 https://doi.org/10.1016/0003-9969(95)00129-8
  38. Iontcheva I, Oppenheim FG, Troxler RF. Human salivary mucin MG1 selectively forms heterotypic complexes with amylase, proline-rich proteins, statherin, and histatins. J Dent Res 1997;76:734-743 https://doi.org/10.1177/00220345970760030501
  39. Ihalin R, Loimaranta V, Tenovuo J. Origin, structure, and biological activities of peroxidases in human saliva. Arch Biochem Biophys 2006;445:261-268 https://doi.org/10.1016/j.abb.2005.07.004
  40. Shannon IL, McCrary BR, Starcke EN. A saliva substitute for use by xerostomic patients undergoing radiotherapy to the head and neck. Oral Surg 1977;44:656-661 https://doi.org/10.1016/0030-4220(77)90373-5
  41. Hannig C, Attin T, Hannig M, Henze E, Brinkmann K, Zech R. Immobilisation and activity of human ${\alpha}$-amylase in the acquired enamel pellicle. Arch Oral Biol 2004;49:469-475 https://doi.org/10.1016/j.archoralbio.2004.01.005
  42. Christersson CE, Lindh L, Arnebrant T. Film-forming properties and viscosities of saliva substitutes and human whole saliva. Eur J Oral Sci 2000;108:418-425 https://doi.org/10.1034/j.1600-0722.2000.108005418.x
  43. Jiang W, Gupta D, Gallagher D, Davis S, Bhavanandan VP. The central domain of bovine submaxillary mucin consists of over 50 tandem repeats of 329 amino acids. Chromosomal localization of the BSM1 gene and relations to ovine and porcine counterparts. Eur J Biochem 2000;267:2208-2217 https://doi.org/10.1046/j.1432-1327.2000.01225.x
  44. Tenovuo J. Clinical applications of antimicrobial host proteins lactoperoxidase, lysozyme and lactoferrin in xerostomia: efficacy and safety. Oral Dis 2002;8:23-29 https://doi.org/10.1034/j.1601-0825.2002.1o781.x
  45. Sandwick RK, Schray KJ. Conformational states of enzymes bound to surfaces. J Coll Interface Sci 1988;121:1-12 https://doi.org/10.1016/0021-9797(88)90402-X
  46. McGaughey C, Stowell EC. The adsorption of human salivary proteins and porcine submaxillary mucin by hydroxyapatite. Arch Oral Biol 1967;12:815-828 https://doi.org/10.1016/0003-9969(67)90104-5
  47. Bruno LS, Li X, Wang L, Soares RV, Siqueira CC, Oppenheim FG, Troxler RF, Offner GD. Two-hybrid analysis of human salivary mucin MUC7 ineractions. Biochimica et Biophysica Acta 2005;1746:65-72 https://doi.org/10.1016/j.bbamcr.2005.08.007
  48. Shomers JP, Tabak LA, Levine MJ, Mandel ID, Ellison SA. The isolation of a family of cysteine- containing phosphoproteins from humansubmandibular-sublingual saliva. J Dent Res 1982;61:973-977 https://doi.org/10.1177/00220345820610081101
  49. Nordman H, Davies JR, Herrmann A, Karlsson NG, Hansson GC, Carlstedt I. Mucus glycoproteins from pig gastric mucosa: identification of different mucin populations from the surface epithelium. Biochem J 1997;326:903-910 https://doi.org/10.1042/bj3260903
  50. Kousvelari EE, Baratz RS, Burke B, Oppenheim FG. Immunochemical identification and determination of proline-rich proteins in salivary secretions, enamel pellicle, and glandular tissue specimens. J Dent Res 1980;59:1430-1438 https://doi.org/10.1177/00220345800590081201
  51. Pruitt KM, Adamson M. Enzyme activity of salivary lactoperoxidase adsorbed to human enamel. Infect Immun 1977;17:112-116
  52. Kho HS, Vacca Smith AM, Koo H, Scott-Anne K, Bowen WH. Interactions of Streptococcus mutans glucosyltransferase B with lysozyme in solution and on the surface of hydroxyapatite. Caries Res 2005;39:411-416 https://doi.org/10.1159/000086849
  53. Mnsson-Rahemtulla B, Baldone DC, Pruitt KM, Rahemtulla F. Specific assays for peroxidases in human saliva. Arch Oral Biol 1986;31:661-668 https://doi.org/10.1016/0003-9969(86)90095-6