Characterization of a serine protease homologous to house dust mite group 3 allergens from the scabies mite sarcoptes scabiei

Simone Alexandra Beckham, Sarah Elizabeth Boyd, Simone Reynolds, Charlene Willis, Masego Johnstone, Angela Mika, Pavla Simerska, Lakshmi C Wijeyewickrema, Alexander Ian Smith, David J Kemp, Robert Neil Pike, Katja Fischer

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32 Citations (Scopus)

Abstract

The scabies mite, Sarcoptes scabiei var. hominis, infests human skin, causing allergic reactions and facilitating bacterial infection by Streptococcus sp., with serious consequences such as rheumatic fever and rheumatic heart disease. To identify a possible drug target or vaccine candidate protein, we searched for homologues of the group 3 allergen of house dust mites, which we subsequently identified in a cDNA library. The native protein, designated Sar s 3, was shown to be present in the mite gut and excreted in fecal pellets into mite burrows within the upper epidermis. The substrate specificity of proteolytically active recombinant rSar s 3 was elucidated by screening a bacteriophage library. A preference for substrates containing a RSG/A sequence at the P1-P2 positions was revealed. A series of peptides synthesized as internally quenched fluorescent substrates validated the phage display data and HPLC/MS analysis of the preferred cleaved substrate confirmed the predicted cleavage site. Searches of the human proteome using sequence data from the phage display allowed the in silico prediction of putative physiological substrates. Amongst these were numerous epidermal proteins, with filaggrin being a particularly likely candidate substrate. We showed that recombinant rSar s 3 cleaves human filaggrin in vitro and obtained immunohistological evidence that the filaggrin protein is ingested by the mite. This is the first report elucidating the substrate specificity of Sar s 3 and its potential role in scabies mite biology.
Original languageEnglish
Pages (from-to)34413 - 34422
Number of pages10
JournalThe Journal of Biological Chemistry
Volume284
Issue number49
Publication statusPublished - 2009

Cite this

Beckham, S. A., Boyd, S. E., Reynolds, S., Willis, C., Johnstone, M., Mika, A., Simerska, P., Wijeyewickrema, L. C., Smith, A. I., Kemp, D. J., Pike, R. N., & Fischer, K. (2009). Characterization of a serine protease homologous to house dust mite group 3 allergens from the scabies mite sarcoptes scabiei. The Journal of Biological Chemistry, 284(49), 34413 - 34422.