TY - JOUR
T1 - Total and Lectin-Binding Proteome of Spherulin from Coccidioides posadasii
AU - Grys, Thomas E.
AU - Kaushal, Setu
AU - Chowdhury, Yasmynn
AU - Dasari, Surendra
AU - Mitchell, Natalie Michelle
AU - Magee, Dewey
AU - Blair, Janis E.
AU - Colby, Thomas V.
AU - Lake, Douglas
N1 - Publisher Copyright:
© 2016 American Chemical Society.
PY - 2016/10/7
Y1 - 2016/10/7
N2 - 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.
AB - 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.
KW - coccidioidomycosis
KW - fungal infection
KW - proteome of Coccidioides posadasii
UR - http://www.scopus.com/inward/record.url?scp=84990866223&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84990866223&partnerID=8YFLogxK
U2 - 10.1021/acs.jproteome.5b01054
DO - 10.1021/acs.jproteome.5b01054
M3 - Article
C2 - 27546806
AN - SCOPUS:84990866223
SN - 1535-3893
VL - 15
SP - 3463
EP - 3472
JO - Journal of Proteome Research
JF - Journal of Proteome Research
IS - 10
ER -