Abstract
Cryptococcus neoformans var. neoformans is an important fungal pathogen. The capsule is a well established virulence factor and a target site for diagnostic tests. The CPU gene is required for capsular formation and virulence. The protein product Cpll has been proposed to be a secreted protein, but the characteristics of this protein have not been reported. Here we sought to characterize Cpll. Phylogenetic analysis showed that the Cpll of C. neoformans var. neoformans and lite Cpll orthologs identified in C. neoformans var. grubii and C. gattii formed a distinct cluster among related fungi; while the putative ortholog found in Trichosporon asahii was distantly related to the Cryptococcus cluster. We expressed Cpll abundantly as a secreted His-tagged protein in Pichia pastoris. The protein was used to immunize guinea pigs and rabbits for high titer mono-specific polyclonal antibody that was shown to be highly specific against the cell wall of C. neoformans var. neoformans and did not cross react with C. gattii, T. asahii, Aspergillus spp., Candida spp. and Penicillium spp. Using the anti-Cpll antibody, we detected Cpll protein in the fresh culture supernatant of C. neoformans var. neoformans and we showed by immunostaining that the Cpll protein was located on the surface. The Cpll protein is a specific surface protein of C. neoformans var. neoformans.
| Original language | English |
|---|---|
| Pages (from-to) | 39-45 |
| Number of pages | 7 |
| Journal | Mycologia |
| Volume | 107 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 1 2015 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2015 by The Mycological Society of America.
ASJC Scopus Subject Areas
- Ecology, Evolution, Behavior and Systematics
- Physiology
- Molecular Biology
- Genetics
- Cell Biology
Keywords
- Capsule
- CLPI
- Cryptococcus
- Fungus
- Immunostaining