title: Total and Lectin-Binding Proteome of Spherulin from Coccidioides posadasii.
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.
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic , Phoenix, Arizona 85054, United States.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Grys
authors.affiliation: School of Life Sciences, Mayo Clinic Collaborative Research Building, Arizona State University , Scottsdale, Arizona 85259, United States.
authors.forename: Setu
authors.initials: S
authors.lastname: Kaushal
authors.affiliation: School of Life Sciences, Mayo Clinic Collaborative Research Building, Arizona State University , Scottsdale, Arizona 85259, United States.
authors.forename: Yasmynn
authors.initials: Y
authors.lastname: Chowdhury
authors.affiliation: Proteomics Core, Mayo Clinic , Rochester, Minnesota 55902, United States.
authors.forename: Surendra
authors.initials: S
authors.lastname: Dasari
authors.affiliation: School of Life Sciences, Mayo Clinic Collaborative Research Building, Arizona State University , Scottsdale, Arizona 85259, United States.
authors.forename: Natalie Michelle
authors.initials: NM
authors.lastname: Mitchell
authors.affiliation: Center for Personalized Diagnostics, Biodesign Institute, Arizona State University , Tempe, Arizona 85281, United States.
authors.forename: D Mitchell
authors.initials: DM
authors.lastname: Magee
authors.affiliation: Division of Infectious Diseases, Mayo Clinic , Phoenix, Arizona 85054, United States.
authors.forename: Janis E
authors.initials: JE
authors.lastname: Blair
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic , Phoenix, Arizona 85054, United States.
authors.forename: Thomas V
authors.initials: TV
authors.lastname: Colby
authors.affiliation: School of Life Sciences, Mayo Clinic Collaborative Research Building, Arizona State University , Scottsdale, Arizona 85259, United States.
authors.forename: Douglas F
authors.initials: DF
authors.lastname: Lake
mesh_terms: D002846:Chromatography, Affinity; D003045:Coccidioides / Q000737:chemistry*; D003046:Coccidioidin / Q000737:chemistry*; D003047:Coccidioidomycosis / Q000175:diagnosis / Q000473:pathology; D005656:Fungal Proteins / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007150:Immunohistochemistry; D037102:Lectins / Q000378:metabolism*; D008168:Lung / Q000473:pathology; D011485:Protein Binding; D020543:Proteome / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D006023:Glycoproteins; D037102:Lectins; D020543:Proteome; C009303:spherulin; D003046:Coccidioidin
keywords: coccidioidomycosis; fungal infection; proteome of Coccidioides posadasii