Skip to main content

Advertisement

SpringerLink
Log in

Characterization of the ovine ortholog of secretory leukoprotease inhibitor

  • Published:
Mammalian Genome Aims and scope Submit manuscript

Abstract

There is great interest in the use of the sheep as a model for the investigation of inflammation in the lung. The serine antiproteases secretory leukoprotease inhibitor (SLPI) and elafin are important “alarm antiproteases” in the lung and have potentially important roles in the innate immune response. SLPI was first characterized in man and subsequently in murine, porcine, and rat tissues. Here we present the first data concerning the gene and cDNA sequence encoding for the ovine ortholog of SLPI, a protein of 132 amino acids with 66% sequence identity at the amino acid level with human SLPI. A 24-amino-acid signal sequence signifies that, like the other mammalian ortholoags, ovine SLPI is a secreted protein. Tissue distribution of expression is demonstrated by reverse transcription polymerase chain reaction (RT-PCR) and shows features similar to SLPI expression in other mammals, specifically at mucosal surfaces such as the upper respiratory and intestinal tracts, and also the skin, liver, and kidney. This distribution lends credence to SLPI having important roles in innate immunity. We have also cloned the ovine SLPI cDNA into an expression vector and expressed the ovine SLPI protein in vitro. This has enabled us to demonstrate that ovine SLPI is correctly processed (Western blot analysis and SELDI-TOF mass spectrometry analysis) and has biological antihuman neutrophil elastase activity. In summary, the ovine ortholog of SLPI shows similarities to other members of the SLPI family and has all the features of a modulator of innate immunity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2 
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Boudier C, Bieth JG (1992) The proteinase:mucus proteinase inhibitor stoichiometry. J Biol Chem 267:4370-4375

    PubMed  Google Scholar 

  • Brown TI, Mistry R, Collie D, Tate S, Sallenave J-M (2004) Trappin ovine molecule (TOM), the ovine ortholog of elafin, is an acute phase reactant in the lung. Physiol Genomics 19:11-21

    Article  PubMed  Google Scholar 

  • Eisenberg SP, Hale KK, Heimdal P, Thompson RC (1990) Location of the protease-inhibitory region of secretory leukoprotease inhibitor. J Biol Chem 265:7976-7981

    PubMed  Google Scholar 

  • Farmer SJ, Fliss AE, Simmen RC (1990) Complementary DNA cloning and regulation of expression of the messenger RNA encoding a pregnanacy-associated porcine uterine protein related to human antileukoproteinase. Mol Endocrinol 4:1095-1104

    PubMed  Google Scholar 

  • Franken C, Meijer CJ, Dijkman JH (1989) Tissue distribution of antileukoprotease and lysozyme in humans. J Histochem Cytochem 37:493-498

    PubMed  Google Scholar 

  • Gross P, Pfitzer E, Tolker E, Babyak M, Kaschak M (1965) Experimental emphysema: its production with papain in normal and silicotic rats. Arch Environ Health 11:50–58

    PubMed  Google Scholar 

  • Grutter MG, Fendrich G, Huber R, Bode W (1988) The 2.5 ( X-ray crystal structure of the acid-stable proteinase inhibitor from human mucous secretions analysed in its complex with bovine alpha-chymotrypsin. EMBO J 7:345-351

    PubMed  Google Scholar 

  • Heinzel R, Appelhans H, Gassen G, Seemuller U, Machleidt W, et al. (1986) Molecular cloning and expression of cDNA for human antileukoprotease from cervix uterus. Eur J Biochem 160:61-67

    Article  PubMed  Google Scholar 

  • Hiemstra PS, Maassen RJ, Stolk J, Heinzel-Wieland R, Steffens GJ, et al. (1996) Antibacterial activity of antileukoprotease. Infect Immun 64:4520-4524

    PubMed  Google Scholar 

  • Jin FY, Nathan C, Radzioch D, Ding A (1997) Secretory leukoprotease inhibitor: a macrophage product induced by and antagonistic to bacterial lipopolysaccharide. 88:417-426

  • Laurell CB, Erikkson S (1963) Electrophoretic alpha-1-globulin pattern of serum in alpha-1-antitrypsin deficiency. Scand J Clin Lab Invest 15:132–140

    Google Scholar 

  • Nara K, Ito S, Ito T, Suzuki Y, Ghoneim MA, et al. (1994) Elastase inhibitor elafin is a new type of proteinase inhibitor which has a transglutaminase-mediated anchoring sequence termed “cementoin.” J Biochem (Tokyo) 115:441-448

    Google Scholar 

  • Ohlsson S, Ljungkrantz I, Ohlsson K, Segelmark M, Wieslander J (2001) Novel distribution of the secretory leukocyte proteinase inhibitor in kidney. Mediators Inflamm 10:347-350

    Article  PubMed  Google Scholar 

  • Rudolphus A, Stolk J, Dijkman JH, Kramps JA (1993) Inhibition of lipopolysaccharide-induced pulmonary emphysema by intratracheally instilled recombinant secretory leukocyte proteinase inhibitor. Am Rev Respir Dis 147:442-447

    PubMed  Google Scholar 

  • Sallenave J-M, Ryle AP (1991) Purification and characterization of elastase-specific inhibitor. Sequence homology with mucuc proteinase inhibitor. Biol Chem Hoppe Seyler 372:13-21

    PubMed  Google Scholar 

  • Sallenave J-M, Silva A, Marsden ME, Ryle A (1993) Secretion of mucus proteinase inhibitor and elafin by Clara cell and type II pneumocyte cell lines. Am J Respir Cell Mol Biol 8:126-133

    PubMed  Google Scholar 

  • Schalkwijk J, Wiedow O, Hirose S (1999) The trappin gene family: proteins defined by an N-terminal transglutaminase substrate domain and a C-terminal four-disulphide core. Biochem J 340:569-577

    Article  PubMed  Google Scholar 

  • Seemuller U, Arnhold M, Fritz H, Wiedenmann K, Machleidt W, et al. (1986) The acid-stable proteinase inhibitor of human mucus secretions (HUSI-I, antileukoprotease). Complete amino acid sequence as revealed by protein and cDNA sequencing and structural homology to whey proteins and Red Sea turtle proteinase inhibitor. FEBS Lett 199:43-48

    Article  PubMed  Google Scholar 

  • Simpson AJ, Maxwell AI, Govan JR, Haslett C, Sallenave J-M (1999) Elafin (elastase-specific inhibitor) has anti-microbial activity against gram-positive and gram-negative respiratory pathogens. FEBS Lett 452:309-313

    Article  PubMed  Google Scholar 

  • Song X, Zeng L, Jin W, Thompson J, Mizel DE, et al. (1999) Secretory leukocyte proteinase inhibitor suppresses the inflammation and joint damage of bacterial cell wall-induced arthritis. J Exp Med 190:535-542

    Article  PubMed  Google Scholar 

  • Sponer M, Nick HP, Schnebli HP (1991) Different susceptibility of elastase inhibitors to inactivation by proteinases from Staphylococcus aureus and Pseudomonas aeruginosa. Biol Chem Hoppe Seyler 372:963-970

    PubMed  Google Scholar 

  • Stetler G, Brewer MT, Thompson RC (1986) Isolation and sequence of a gene encoding a potent inhibitor of leukocyte proteases. Nucleic Acids Res 14:7883-7896

    PubMed  Google Scholar 

  • Stolk J, Rudolphus A, Kramps JA (1991) Lipopolysaccharide-induced alveolar wall destruction in the hamster is inhibited by intratracheal treatment with r-secretory leukoprotease inhibitor. Ann N Y Acad Sci 624:350-352

    PubMed  Google Scholar 

  • Taggart CC, Greene CM, McElvaney NG, O’Neill S (2002) Secretory leukoprotease inhibitor prevents lipopolysaccharide-induced IkappaBalpha degradation without affecting phosphorylation or ubiquitination. J Biol Chem 277:33648-33653

    Article  PubMed  Google Scholar 

  • Thompson RC, Ohlsson K (1986) Isolation, properties and complete amino acid sequence of human secretory leukoprotease inhibitor, a potent inhibitor of leukocyte elastase. Proc Natl Acad Sci U S A 83:6692-6696

    PubMed  Google Scholar 

  • Tomee JF, Hiemstra PS, Heinzel-Wieland R, Kauffman HF (1997) Antileukoprotease: an endogenous protein in the innate mucosal defense against fungi. J Infect Dis 176:740-747

    PubMed  Google Scholar 

  • Tsunemi M, Matsuura Y, Sakakibara S, Katsube Y (1996) Crystal structure of an elastase-specific inhibitor elafin complexed with porcine pancreatic elastase determined at 1.9 ( resolution. Biochemistry 35:11570-11576

    Article  PubMed  Google Scholar 

  • Zitnik RJ, Zhang J, Kashem MA, Kohno T, Lyons DE, et al. (1997) The cloning and characterization of a murine secretory leukocyte proteinase inhibitor cDNA. Biochem Biophys Res Commun 232:687-697

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to the Wellcome Trust for funding this work (Clinical Research Training Award in Veterinary Medicine for T. Brown) and to M. Marsden for expert technical assistance. The MRC and the Cystic Fibrosis Trust are also acknowledged for funding the SELDI-related experimental work. We also thank Prof. K. Johnson (Molecular Genetics, Anderson College, Institute of Biomedical Sciences, University of Glasgow, UK) and Dr. T. D. Tetley (Lung Cell Biology, National Heart and Lung Institute, Imperial College, London, UK) for their help and input in this project.

Author information

Authors and Affiliations

  1. Rayne Laboratory, Respiratory Medicine Unit, MRC Centre for Inflammation Research, University of Edinburgh, Edinburgh, Scotland, EHS 9AG, UK

    Thomas I. Brown & Jean-Michel Sallenave

  2. Wellcome Trust Centre for Research in Comparative Respiratory Medicine, The University of Edinburgh, Easter Bush Veterinary Centre, Roslin Midlothian, Scotland, UK

    Thomas I. Brown & David D. Collie

  3. Molecular Genetics, Anderson College, Institute of Biomedical Sciences, University of Glasgow, Glasgow, Scotland, UK

    Rohit Mistry

  4. Molecular Medicine Centre, Western General Hospital, Edinburgh, Scotland, UK

    Robert Gray & Margaret Imrie

Authors
  1. Thomas I. Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rohit Mistry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robert Gray
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Margaret Imrie
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David D. Collie
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jean-Michel Sallenave
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jean-Michel Sallenave.

Additional information

Abbreviations:SLPI: secretory leukoprotease inhibitor; BAC: bacterial artificial chromosome; RACE: rapid amplification of cDNA ends; WAP: whey acidic protein; REST: rapidly evolving seminal vesicle transcribed; TOM: trappin ovine molecule; HNE: human neutrophil elastase; EIC: elastase inhibitory capacity; PAGE: polyacrylamide gel electrophoresis; UTR: untranslated region ; SELDI-TOF: surface-enhanced laser desorption/ionization time of flight.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Brown, T.I., Mistry, R., Gray, R. et al. Characterization of the ovine ortholog of secretory leukoprotease inhibitor. Mamm Genome 16, 621–630 (2005). https://doi.org/10.1007/s00335-005-0030-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-005-0030-2

Keywords

Search

Navigation