title: Two-dimensional gel proteomic analysis of Dermatophagoides farinae feces.
abstract: Dermatophagoides farinae fecal allergens are a major source of immunogens in home environments; however, as the source of mite fecal allergen is considered spent growth medium extract that can only mimic the pure fecal extract. In this study, we prepared and using proteomic methods analyzed a D. farinae fecal extract for the first time. The preparation approach used D. farinae feces that were produced within 8 weeks of initiating cultivation in minimized growth media. The feces were collected via adhesion to the tissue culture flask surfaces after removing the SGM and mites. This study contains in-depth proteomic mapping of the allergenic isoforms from the D. farinae fecal extract. Despite extensive analysis, MALDI TOF/TOF spectrometry showed that only six proteins/allergens, Der f1, Der f2, Der f3, Der f6, Der f15 and ferritin, originated from D. farinae. No other analyzed proteins were exactly assigned to Dermatophagoides or to similar invertebrate species by sequence similarity. The remaining proteins were assigned mostly to yeasts or cereals (originally dietary proteins); however, many of the proteins were not successfully identified in the current NCBInr. The numerous dietary proteins identified in the feces suggest that these proteins remained highly stable after passing through the gut. Isoforms of the allergens Der f1, Der f3 and Der f15 were identified in more MWs indicating the presence of zymogens and active-enzyme forms. The identified fecal allergens accumulate in the environment during the life of the mite and represent quantitatively greater amounts of mite immunogens than those that were missed in the 2D-E. The results contribute to our understanding of D. farinae digestive physiology with regard to the enzymes/proteins present in the feces.
authors.affiliation: Biologically Active Substances in Crop Protection, Laboratory of Proteomics, Crop Research Institute, Drnovska 507/73, 16106, Praha 6-Ruzyne, Czech Republic, arachnid@centrum.cz.
authors.forename: Tomas
authors.initials: T
authors.lastname: Erban
authors.forename: Jan
authors.initials: J
authors.lastname: Hubert
mesh_terms: D000818:Animals; D039741:Antigens, Dermatophagoides / Q000737:chemistry*; D040001:Dermatophagoides farinae / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005243:Feces / Q000737:chemistry*; D020033:Protein Isoforms; D040901:Proteomics / Q000379:methods*
chemical_list: D039741:Antigens, Dermatophagoides; D020033:Protein Isoforms
references.citation: J Allergy Clin Immunol. 1998 Aug;102(2):295-303
references.pmid: 9723675
references.citation: Allergy. 2002 Aug;57(8):729-31
references.pmid: 12121193
references.citation: J Mol Biol. 2009 Feb 20;386(2):520-30
references.pmid: 19136006
references.citation: Chang Gung Med J. 2004 Aug;27(8):563-9
references.pmid: 15553602
references.citation: Clin Exp Allergy. 2002 Sep;32(9):1341-7
references.pmid: 12220473
references.citation: J Allergy Clin Immunol. 2003 Jul;112(1):79-86
references.pmid: 12847483
references.citation: J Med Entomol. 1997 Nov;34(6):684-9
references.pmid: 9439123
references.citation: J Med Entomol. 1998 Nov;35(6):962-6
references.pmid: 9835687
references.citation: J Med Entomol. 1996 Mar;33(2):257-60
references.pmid: 8742531
references.citation: Int Arch Allergy Appl Immunol. 1985;76(1):58-67
references.pmid: 3967933
references.citation: J Investig Allergol Clin Immunol. 1998 Jul-Aug;8(4):201-6
references.pmid: 9777533
references.citation: Clin Exp Allergy. 1995 Aug;25(8):765-70
references.pmid: 7584689
references.citation: Mol Cell Proteomics. 2013 Jul;12(7):1818-28
references.pmid: 23481662
references.citation: J Allergy Clin Immunol. 2007 Jul;120(1):105-12
references.pmid: 17445876
references.citation: Med Vet Entomol. 2011 Jun;25(2):209-16
references.pmid: 21198710
references.citation: Allergy. 2004 Jun;59(6):571-4
references.pmid: 15147439
references.citation: Int Arch Allergy Appl Immunol. 1991;95(2-3):248-56
references.pmid: 1718911
references.citation: Arch Insect Biochem Physiol. 2010 Nov;75(3):187-206
references.pmid: 20936642
references.citation: Immunology. 1992 Jan;75(1):29-35
references.pmid: 1537598
references.citation: J Allergy Clin Immunol. 1989 Jun;83(6):1055-67
references.pmid: 2732406
references.citation: Trends Mol Med. 2010 Jul;16(7):321-8
references.pmid: 20605742
references.citation: Clin Exp Allergy. 2000 Sep;30(9):1293-7
references.pmid: 10971476
references.citation: Allergy. 2010 Feb;65(2):184-90
references.pmid: 19796217
references.citation: J Immunol. 2005 Sep 15;175(6):3835-45
references.pmid: 16148130
references.citation: Arch Insect Biochem Physiol. 2009 Jul;71(3):139-58
references.pmid: 19480003
references.citation: Clin Exp Allergy. 2005 Oct;35(10):1384-91
references.pmid: 16238800
references.citation: Vet Immunol Immunopathol. 2001 Feb 10;78(3-4):231-47
references.pmid: 11292526
references.citation: Nature. 1981 Feb 12;289(5798):592-3
references.pmid: 7464922
references.citation: J Insect Sci. 2010;10:42
references.pmid: 20572792
references.citation: Int Arch Allergy Immunol. 2006;140(4):295-305
references.pmid: 16741365
references.citation: Exp Appl Acarol. 2008 Mar;44(3):199-212
references.pmid: 18357505
references.citation: J Allergy Clin Immunol. 2006 Mar;117(3):571-6
references.pmid: 16522455
references.citation: Immunol Rev. 2011 Jul;242(1):51-68
references.pmid: 21682738
references.citation: Allergy. 2008 Jun;63(6):758-67
references.pmid: 18445190
references.citation: J Immunol. 2013 Apr 1;190(7):3059-67
references.pmid: 23460742
references.citation: J Med Entomol. 2010 Nov;47(6):1062-70
references.pmid: 21175054
references.citation: J Allergy Clin Immunol. 1998 Oct;102(4 Pt 1):631-6
references.pmid: 9802372
references.citation: Biochem Biophys Res Commun. 2005 Mar 25;328(4):944-52
references.pmid: 15707969
references.citation: Exp Appl Acarol. 1992 Nov;16(1-2):165-80
references.pmid: 1283568
references.citation: J Allergy Clin Immunol. 1989 Feb;83(2 Pt 1):416-27
references.pmid: 2645343
references.citation: Int Arch Allergy Immunol. 2002 Sep;129(1):1-18
references.pmid: 12372994
references.citation: J Med Entomol. 1991 Jul;28(4):487-91
references.pmid: 1941906
references.citation: Ann Allergy Asthma Immunol. 2002 Jul;89(1):34-7
references.pmid: 12141717
references.citation: J Allergy Clin Immunol. 1996 Oct;98(4):739-47
references.pmid: 8876548
references.citation: Allergol Immunopathol (Madr). 1997 May-Jun;25(3):113-7
references.pmid: 9208047
references.citation: Allergy Asthma Immunol Res. 2012 Nov;4(6):346-50
references.pmid: 23115731
references.citation: Clin Exp Allergy. 1997 Feb;27(2):201-7
references.pmid: 9061221