Molecular cloning and expression of a cDNA encoding a Coccidioides posadasii Cu,Zn superoxide dismutase identified by proteomic analysis of the coccidioidal T27K vaccine.
Previous studies have demonstrated that the coccidioidal T27K vaccine preparation is protective in mice against respiratory challenge using Coccidioides posadasii (C. posadasii) arthroconidia. Proteomic methods have been employed to define the molecular components within the vaccine. This method has led to the identification of novel and previously uncharacterized coccidioidal proteins including a Cu,Zn superoxide dismutase. A two-dimensional gel of the T27K vaccine was run and spots were excised for mass spectrometric analysis. One peptide was obtained from the T27K gel that matched a TIGR C. posadasii 2.0 gene index tentative consensus sequence, TC1072, which is similar to fungal Cu,Zn superoxide dismutase. Activity assays performed with native PAGE gels of the T27K vaccine showed that the vaccine contains superoxide dismutase. The cDNA encoding the enzyme has been cloned and sequenced and expressed as a recombinant protein.
Lunetta
Jennine M
JM
Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis, California 95616, USA. jmlunetta@ucdavis.edu
Simmons
Keira A
KA
Johnson
Suzanne M
SM
Pappagianis
Demosthenes
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003045:Coccidioides / Q000235:genetics*; D018076:DNA, Complementary / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005657:Fungal Vaccines / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000235:genetics* / Q000502:physiology*
D018076:DNA, Complementary; D005657:Fungal Vaccines; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase