Abstract

Coccidioides is a virulent dimorphic fungus that causes coccidioidomycosis (valley fever) in mammals, including humans. Although the genome has been sequenced, a proteomic analysis does not exist. To address this gap in proteomic knowledge, we generated the proteome of spherulin (a well-studied lysate of fungal spherules) and identified 1390 proteins. Some of the proteins included glycosylation enzymes, which led us to hypothesize that fungal glycosylation patterns may be different from those of mammals and could be exploited to detect Coccidioides in tissues. We performed lectin-based immunohistochemistry on formalin-fixed paraffin-embedded human patients' lung tissues. GSL-II (Griffonia simplificonia lectin II) and sWGA (succinylated wheat germ agglutinin) lectins bound specifically to endospores and spherules in infected lungs. To identify lectin-binding glycoproteins in spherulin, we performed lectin-affinity chromatography, followed by LC-MS/MS. A total of 195 glycoproteins from spherulin bound to GSL-II, 224 glycoproteins bound to sWGA, and 145 glycoproteins bound to both lectins. This is the first report of the specific reactivity of GSL-II and sWGA lectins to Coccidioides endospores and spherules in infected human tissues and the first listing of the Coccidioidal proteome from spherulin using sequences present in three Coccidioides databases: RefSeq, SwissProt, and The Broad Institute's Coccidioides Genome project.

Original languageEnglish (US)
Pages (from-to)3463-3472
Number of pages10
JournalJournal of Proteome Research
Volume15
Issue number10
DOIs
StatePublished - Oct 7 2016

Fingerprint

Coccidioides
Proteome
Lectins
Griffonia
Glycoproteins
Coccidioidomycosis
Wheat Germ Agglutinins
Glycosylation
Proteomics
Mammals
Tissue
Genome
Lung
Protein Databases
Genes
Affinity chromatography
spherulin
Affinity Chromatography
Paraffin
Formaldehyde

Keywords

  • coccidioidomycosis
  • fungal infection
  • proteome of Coccidioides posadasii

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry

Cite this

Total and Lectin-Binding Proteome of Spherulin from Coccidioides posadasii. / Grys, Thomas E.; Kaushal, Setu; Chowdhury, Yasmynn; Dasari, Surendra; Mitchell, Natalie Michelle; Magee, Dewey; Blair, Janis E.; Colby, Thomas V.; Lake, Douglas.

In: Journal of Proteome Research, Vol. 15, No. 10, 07.10.2016, p. 3463-3472.

Research output: Contribution to journalArticle

Grys, TE, Kaushal, S, Chowdhury, Y, Dasari, S, Mitchell, NM, Magee, D, Blair, JE, Colby, TV & Lake, D 2016, 'Total and Lectin-Binding Proteome of Spherulin from Coccidioides posadasii', Journal of Proteome Research, vol. 15, no. 10, pp. 3463-3472. https://doi.org/10.1021/acs.jproteome.5b01054
Grys, Thomas E. ; Kaushal, Setu ; Chowdhury, Yasmynn ; Dasari, Surendra ; Mitchell, Natalie Michelle ; Magee, Dewey ; Blair, Janis E. ; Colby, Thomas V. ; Lake, Douglas. / Total and Lectin-Binding Proteome of Spherulin from Coccidioides posadasii. In: Journal of Proteome Research. 2016 ; Vol. 15, No. 10. pp. 3463-3472.
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