Domain 15 of the serine proteinase inhibitor LEKTI blocks HIV infection in vitro

  • David Palesch Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany
  • Edina Varga Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany
  • Ute C. Marx Lehrstuhl Biopolymere, University of Bayreuth, Bayreuth, Germany
  • Klaus Vitzithum Lehrstuhl Biopolymere, University of Bayreuth, Bayreuth, Germany
  • Peter Kreutzmann IPF PharmaCeuticals GmbH, Hanover, Germany
  • Wolf-Georg Forssmann Clinic of Immunology and Rheumatology, Peptide Research Group, Hanover Medical School, Hanover, Germany
  • Jan Münch Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany
  • Hans-Jürgen Mägert Institute of Biotechnology, Anhalt University of Applied Sciences, Köthen, Germany
Abstract viewed: 1070 times
PDF downloaded: 303 times

Abstract

Background: Lympho-epithelial Kazal-type-related inhibitor (LEKTI) is a 15-domain serine proteinase inhibitor, parts of which have first been isolated from human blood filtrate. It is encoded by the gene SPINK5. In the past, different groups reported antiviral activities of certain serine proteinase inhibitors, such as mucous proteinase inhibitor and alpha1-proteinase inhibitor. The purpose of this study was to test two representative domains of the proteinase inhibitor LEKTI for anti-HIV activities.

Methods: LEKTI domains 6 and 15 were recombinantly produced in E.coli. To test their inhibitory activity against HIV infection, the reporter cell line P4-R5 MAGI carrying an HIV-inducible reporter gene was infected by a CCR5-tropic HIV strain in the presence of different inhibitor concentrations. After three days, infection rates were determined by quantifying ß-galactosidase activities using the Galacto-Light Plus™ ß-Galactosidase Reporter Gene Assay.

Results: In contrast to LEKTI domain 6, LEKTI domain 15 suppressed HIV-induced reporter gene activities with an IC50 value of approximately 29 µM.

Conclusion: LEKTI domain 15 represents an inhibitor of HIV infection. (Med J Indones. 2013;22:131-5. doi: 10.13181/mji.v22i3.580)

Keywords: HIV, inhibition, LEKTI, P4-R5 MAGI

References

  1. Mägert HJ, Kreutzmann P, Ständker L, Walden M, Drögemüller K, Forssmann WG. LEKTI: a multidomain serine proteinase inhibitor with pathophysiological relevance. Int J Biochem Cell Biol. 2002;34(6):573-6. http://dx.doi.org/10.1016/S1357-2725(01)00179-0

  2. Mägert HJ, Kreutzmann P, Drögemüller K, et al. The 15-domain serine proteinase inhibitor LEKTI: biochemical properties, genomic organization, and pathophysiological role. Eur J Med Res. 2002;7(2):49-56.

  3. Bitoun E, Micheloni A, Lamant L, et al. LEKTI proteolytic processing in human primary keratinocytes, tissue distribution and defective expression in Netherton syndrome. Hum Mol Genet. 2003;12(19):2417-30. http://dx.doi.org/10.1093/hmg/ddg247

  4. Mitsudo K, Jayakumar A, Henderson Y, et al. Inhibition of serine proteinases plasmin, trypsin, subtilisin A, cathepsin G, and elastase by LEKTI: a kinetic analysis. Biochemistry 2003;42(13):3874-81. http://dx.doi.org/10.1021/bi027029v

  5. Egelrud T, Brattsand M, Kreutzmann P, et al. hK5 and hK7, two serine proteinases abundant in human skin, are inhibited by LEKTI domain 6. Br J Dermatol. 2005;153(6):1200-3. http://dx.doi.org/10.1111/j.1365-2133.2005.06834.x

  6. Schechter NM, Choi EJ, Wang ZM, et al. Inhibition of human kallikreins 5 and 7 by the serine protease inhibitor lympho-epithelial Kazal-type inhibitor (LEKTI). Biol Chem. 2005;386(11):1173-84. http://dx.doi.org/10.1515/BC.2005.134

  7. Deraison C, Bonnart C, Lopez F, et al. LEKTI fragments specifically inhibit KLK5, KLK7, and KLK14 and control desquamation through a pH-dependent interaction. Mol Biol Cell. 2007;18(9):3607-19. http://dx.doi.org/10.1091/mbc.E07-02-0124

  8. Chavanas S, Bodemer C, Rochat A, et al. Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome. Nat Genet. 2000;25(2):141-2. http://dx.doi.org/10.1038/75977

  9. Walley AJ, Chavanas S, Moffatt MF, et al. Gene polymorphism in Netherton and common atopic disease. Nat Genet. 2001;29(2):175-8. http://dx.doi.org/10.1038/ng728

  10. Kato A, Fukai K, Oiso N, Hosomi N, Murakami T, Ishii M. Association of SPINK5 gene polymorphisms with atopic dermatitis in the Japanese population. Br J Dermatol. 2003;148(4):665-9. http://dx.doi.org/10.1046/j.1365-2133.2003.05243.x

  11. Liu Q, Xia Y, Zhang W, et al. A functional polymorphism in the SPINK5 gene is associated with asthma in a Chinese Han Population. BMC Med Genet. 2009;10:59. http://dx.doi.org/10.1186/1471-2350-10-59

  12. McNeely TB, Shugars DC, Rosendahl M, Tucker C, Eisenberg SP, Wahl SM. Inhibition of human immunodeficiency virus type 1 infectivity by secretory leukocyte protease inhibitor occurs prior to viral reverse transcription. Blood 1997;90(3):1141-9.

  13. Shapiro L, Pott GB, Ralston AH. Alpha-1-antitrypsin inhibits human immunodeficiency virus type 1. FASEB J. 2001;15(1):115-22. http://dx.doi.org/10.1096/fj.00-0311com

  14. Kreutzmann P, Schulz A, Ständker L, Forssmann WG, Mägert HJ. Recombinant production, purification and biochemical characterization of domain 6 of LEKTI: a temporary Kazal-type-related serine proteinase inhibitor. J Chromatogr B Analyt Technol Biomed Life Sci. 2004;803(1):75-81. http://dx.doi.org/10.1016/j.jchromb.2003.07.016

  15. Vitzithum K, Lauber T, Kreutzmann P, et al. LEKTI domain 15 is a functional Kazal-type proteinase inhibitor. Protein Expr Purif. 2008;57(1):45-56. http://dx.doi.org/10.1016/j.pep.2007.08.012

  16. Wei X, Decker JM, Liu H, et al. Emergence of resistant human immunodeficiency virus type 1 in patients receiving fusion inhibitor (T-20) monotherapy. Antimicrob Agents Chemother 2002;46(6):1896-905. http://dx.doi.org/10.1128/AAC.46.6.1896-1905.2002

  17. Hiebenthal-Millow K, Kirchhoff F. The most frequent naturally occurring length polymorphism in the HIV-1 LTR has little effect on proviral transcription and viral replication. Virology 2002;292(1):169-75. http://dx.doi.org/10.1006/viro.2001.1282

  18. Münch J, Ständker L, Adermann K, et al. Discovery and optimization of a natural HIV-1 entry inhibitor targeting the gp41 fusion peptide. Cell 2007;129(2):263-75. http://dx.doi.org/10.1016/j.cell.2007.02.042

Published
2013-08-22
How to Cite
1.
Palesch D, Varga E, Marx UC, Vitzithum K, Kreutzmann P, Forssmann W-G, Münch J, Mägert H-J. Domain 15 of the serine proteinase inhibitor LEKTI blocks HIV infection <em>in vitro</em&gt;. Med J Indones [Internet]. 2013Aug.22 [cited 2019Nov.22];22(3):131-5. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/580
Section
Basic Medical Research