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Characterisation and expression of SPLUNC2, the human orthologue of rodent parotid secretory protein

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Abstract

We recently described the Palate Lung Nasal Clone (PLUNC) family of proteins as an extended group of proteins expressed in the upper airways, nose and mouth. Little is known about these proteins, but they are secreted into the airway and nasal lining fluids and saliva where, due to their structural similarity with lipopolysaccharide-binding protein and bactericidal/permeability-increasing protein, they may play a role in the innate immune defence. We now describe the generation and characterisation of novel affinity-purified antibodies to SPLUNC2, and use them to determine the expression of this, the major salivary gland PLUNC. Western blotting showed that the antibodies identified a number of distinct protein bands in saliva, whilst immunohistochemical analysis demonstrated protein expression in serous cells of the major salivary glands and in the ductal lumens as well as in cells of minor mucosal glands. Antibodies directed against distinct epitopes of the protein yielded different staining patterns in both minor and major salivary glands. Using RT-PCR of tissues from the oral cavity, coupled with EST analysis, we showed that the gene undergoes alternative splicing using two 5′ non-coding exons, suggesting that the gene is regulated by alternative promoters. Comprehensive RACE analysis using salivary gland RNA as template failed to identify any additional exons. Analysis of saliva showed that SPLUNC2 is subject to N-glycosylation. Thus, our study shows that multiple SPLUNC2 isoforms are found in the oral cavity and suggest that these proteins may be differentially regulated in distinct tissues where they may function in the innate immune response.

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References

  • Ball WD, Mirels L, Hand AR (2003) Psp and Smgb: a model for developmental and functional regulation in the rat major salivary glands. Biochem Soc Trans 31:777–780

    Article  PubMed  CAS  Google Scholar 

  • Bingle CD, Craven CJ (2002) PLUNC: a novel family of candidate host defence proteins expressed in the upper airways and nasopharynx. Hum Mol Genet 11:937–943

    Article  PubMed  CAS  Google Scholar 

  • Bingle CD, Craven CJ (2004) Meet the relatives: a family of BPI and LBP-related proteins. Trends Immunol 25:53–55

    Article  PubMed  CAS  Google Scholar 

  • Bingle CD, Gorr SU (2004) Host defence in oral and airway epithelia: chromosome 20 contributes a new protein family. Int J Biochem Cell Biol 36:2144–2152

    Article  PubMed  CAS  Google Scholar 

  • Bingle CD, Vayarkarnam A (2008) Novel innate immune functions of the whey acidic protein family. Trends Immunol 29:444–453

    Article  PubMed  CAS  Google Scholar 

  • Bingle CD, LeClair EE, Havard S, Bingle L, Gillingham P, Craven CJ (2004) Phylogenetic and evolutionary analysis of the PLUNC gene family. Protein Sci 13:422–430

    Article  PubMed  CAS  Google Scholar 

  • Bingle L, Cross SS, High AS, Wallace WA, Devine DA, Havard S, Campos MA, Bingle CD (2005) SPLUNC1 (PLUNC) is expressed in glandular tissues of the respiratory tract and in cancers with a glandular phenotype. J Path 205:491–497

    Article  PubMed  CAS  Google Scholar 

  • Bingle L, Cross SS, High AS, Wallace WA, Rassl D, Yuan G, Hellstrom I, Campos MA, Bingle CD (2006) WFDC2 (HE4): a potential role in the innate immunity of the oral cavity and respiratory tract and the development of adenocarcinomas of the lung. Respir Res 7:61

    Article  PubMed  Google Scholar 

  • Campos MA, Abreu AR, Nlend MC, Cobas MA, Conner GE, Whitney PL (2004) Purification and characterization of PLUNC from human tracheobronchial secretions. Am J Respir Cell Mol Biol 30:184–192

    Article  PubMed  CAS  Google Scholar 

  • Emes RD, Goodstadt L, Winter EE, Ponting CP (2003) Comparison of the genomes of human and mouse lays the foundation of genome zoology. Hum Mol Genet 12:701–709

    Article  PubMed  CAS  Google Scholar 

  • Geetha C, Venkatesh SG, Dunn BH, Gorr SU (2003) Expression and anti-bacterial activity of human parotid secretory protein (PSP). Biochem Soc Trans 31:815–818

    Article  PubMed  CAS  Google Scholar 

  • Geetha C, Venkatesh SG, Bingle L, Bingle CD, Gorr SU (2005) Design and validation of anti-inflammatory peptides from human parotid secretory protein. J Dent Res 84:149–153

    Article  PubMed  CAS  Google Scholar 

  • Gorr SU, Sotsky JB, Shelar AP, Demuth DR (2008) Design of bacteria-agglutinating peptides derived from parotid secretory protein, a member of the bactericidal/permeability increasing-like protein family. Peptides 29:2118–2127

    Article  PubMed  CAS  Google Scholar 

  • Guo T, Rudnick PA, Wang W, Lee CS, Devoe DL, Balgley BM (2006) Characterization of the human salivary proteome by capillary isoelectric focusing/nanoreversed-phase liquid chromatography coupled with ESI-tandem MS. J Proteome Res 5:1469–1478

    Article  PubMed  CAS  Google Scholar 

  • Haigh B, Hood K, Broadhurst M, Medele S, Callaghan M, Smolenski G, Dines M, Wheeler T (2008) The bovine salivary proteins BSP30a and BSP30b are independently expressed BPI-like proteins with anti-Pseudomonas activity. Mol Immunol 45:1944–1951

    Article  PubMed  CAS  Google Scholar 

  • Helmerhorst EJ, Oppenheim FG (2007) Saliva: a dynamic proteome. J Dent Res 86:680–693

    Article  PubMed  CAS  Google Scholar 

  • Khovidhunkit W, Hachem JP, Medzihradszky KF, Duchateau PN, Shigenaga JK, Moser AH, Movsesyan I, Naya-Vigne J, Kane JP, Feingold KR, Grunfeld C (2005) Parotid secretory protein is an HDL-associated protein with anticandidal activity. Am J Physiol Regul Integr Comp Physiol 288:R1306–R1315

    PubMed  CAS  Google Scholar 

  • Madsen HO, Hjorth JP (1985) Molecular cloning of mouse PSP mRNA. Nucleic Acids Res 13:1–13

    Article  PubMed  CAS  Google Scholar 

  • Mirels L, Ball WD (1992) Neonatal rat submandibular gland protein SMG-A and parotid secretory protein are alternatively regulated members of a salivary protein multigene family. J Biol Chem 267:2679–2687

    PubMed  CAS  Google Scholar 

  • Mirels L, Miranda AJ, Ball WD (1998) Characterization of the rat salivary-gland B1-immunoreactive proteins. Biochem J 330:437–444

    PubMed  CAS  Google Scholar 

  • Plummer C, Douglas CWI (2006) Relationship between the ability of oral streptococci to interact with platelet glycoprotein Ibα and with the salivary low-molecular weight mucin, MG2. FEMS Immunol Med Microbiol 48:390–399

    Article  PubMed  CAS  Google Scholar 

  • Ramachandran P, Boontheung P, Xie Y, Sondej M, Wong DT, Loo JA (2006) Identification of N-linked glycoproteins in human saliva by glycoprotein capture and mass spectrometry. J Proteome Res 5:1493–1503

    Article  PubMed  CAS  Google Scholar 

  • Robinson CP, Bounous DI, Alford CE, Nguyen KT, Nanni JM, Peck AB, Ammon B, Humphreys-Beher MG (1997) PSP expression in murine lacrimal glands and function as a bacteria-binding protein in exocrine secretions. Am J Physiol 272:G863–G871

    PubMed  CAS  Google Scholar 

  • Shaw P, Schibler U (1986) Structure and expression of the parotid secretory protein gene of mouse. J Mol Biol 192:567–576

    Article  PubMed  CAS  Google Scholar 

  • Shiba H, Venkatesh SG, Gorr SU, Barbieri G, Kurihara H, Kinane DF (2005) Parotid secretory protein is expressed and inducible in human gingival keratinocytes. J Periodontal Res 40:153–157

    Article  PubMed  CAS  Google Scholar 

  • Siqueira WL, Zhang W, Helmerhorst EJ, Gygi SP, Oppenheim FG (2007) Identification of protein components in vivo human acquired enamel pellicle using LC-ESI-MS/MS. J Proteome Res 6:2152–2160

    Article  PubMed  CAS  Google Scholar 

  • Siqueira WL, Salih E, Wan DL, Helmerhorst EJ, Oppenheim FG (2008) Proteome of human minor salivary gland secretion. J Dent Res 87:445–450

    Article  PubMed  CAS  Google Scholar 

  • Vargas PA, Speight PM, Bingle CD, Barrett AW, Bingle L (2008) Expression of PLUNC family members in benign and malignant salivary gland tumours. Oral Dis 14:613–619

    Article  PubMed  CAS  Google Scholar 

  • Vitorino R, Lobo MJ, Ferrer-Correira AJ, Dubin JR, Tomer KB, Domingues PM, Amado FM (2004) Identification of human whole saliva protein components using proteomics. Proteomics 4:1109–1115

    Article  PubMed  CAS  Google Scholar 

  • Walz A, Stuhler K, Wattenberg A, Hawranke E, Meyer HE, Schmalz G, Bluggel M, Ruhl S (2006) Proteome analysis of glandular parotid and submandibular–sublingual saliva in comparison to whole human saliva by two-dimensional gel electrophoresis. Proteomics 6:1631–1639

    Article  PubMed  CAS  Google Scholar 

  • Weston WM, LeClair EE, Trzyna W, McHugh KM, Nugent P, Lafferty CM, Ma L, Tuan RS, Greene RM (1999) Differential display identification of plunc, a novel gene expressed in embryonic palate, nasal epithelium, and adult lung. J Biol Chem 274:13698–13703

    Article  PubMed  CAS  Google Scholar 

  • Wheeler TT, Haigh BJ, McCracken JY, Wilkins RJ, Morris CA, Grigor MR (2002) The BSP30 salivary proteins from cattle, LUNX/PLUNC and von Ebner’s minor salivary gland protein are members of the PSP/LBP superfamily of proteins. Biochim Biophys Acta 1579:92–100

    PubMed  CAS  Google Scholar 

  • Wheeler TT, Hood KA, Maqbool N, McEwan JC, Bingle CD, Zhao S (2007) Expansion of the bactericidal/permeability increasing-like (BPI-like) protein locus in cattle. BMC Genomics 8:75

    Article  PubMed  Google Scholar 

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Acknowledgment

This work was funded by The Wellcome Trust and the Sheffield Hospital’s Charitable Trust.

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Authors and Affiliations

  1. Department of Oral and Maxillofacial Pathology, School of Clinical Dentistry, University of Sheffield, Claremont Crescent, Sheffield, S10 2TA, UK

    Lynne Bingle, Hayley Lunn & C. W. Ian Douglas

  2. Academic Unit of Respiratory Medicine, Department of Infection and Immunity, University of Sheffield, Sheffield, UK

    Frances A. Barnes, Maslinda Musa, Steve Webster & Colin D. Bingle

  3. Academic Unit of Pathology, Department of Infection and Immunity, University of Sheffield, Sheffield, UK

    Simon S. Cross

  4. Diagnostic Services Department, Medical and Dental School, University of Leeds, Leeds, UK

    Alec S. High

Authors
  1. Lynne Bingle
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  2. Frances A. Barnes
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  3. Hayley Lunn
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  4. Maslinda Musa
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  5. Steve Webster
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  6. C. W. Ian Douglas
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  7. Simon S. Cross
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  8. Alec S. High
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  9. Colin D. Bingle
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Correspondence to Lynne Bingle.

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Bingle, L., Barnes, F.A., Lunn, H. et al. Characterisation and expression of SPLUNC2, the human orthologue of rodent parotid secretory protein. Histochem Cell Biol 132, 339–349 (2009). https://doi.org/10.1007/s00418-009-0610-4

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  • DOI: https://doi.org/10.1007/s00418-009-0610-4

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