Ubiquitination proteome profiling of Candida albicans

ABSTRACT: Protein extraction The two groups (C, CS) were treated in same way, sample was first grinded with liquid nitrogen, then the powder was transferred to a 5-mL centrifuge tube and sonicated three times on ice using a high intensity ultrasonic processor (Scientz) in lysis buffer (including 1% TritonX-100, 10 mM dithiothreitol, and 1% protease inhibitor cocktail, 50 μM PR-619, 3 μM TSA, 50 mM NAM, 2 mM EDTA, and 1% phosphatase inhibitor for phosphorylation.). An equal volume of Tris-saturated phenol (pH 8.0) was added. Then, the mixture was further vortexed for 5 min. After centrifugation (4 °C, 10 min, 5 000g), the upper phenol phase was transferred to a new centrifuge tube. Proteins were precipitated by adding at least four volumes of ammonium sulfate-saturated methanol and incubated at -20 °C for at least 6 h. After centrifugation at 4 °C for 10 min, the supernatant was discarded. The remaining precipitate was washed with ice-cold methanol, followed by ice-cold acetone for three times. The protein was redissolved in 8 M urea and the protein concentration was determined with BCA kit according to the manufacturer’s instructions.

INSTRUMENT(S):

ORGANISM(S): Candida Albicans (yeast)

TISSUE(S): Eukaryotic Cell, Whole Body

DISEASE(S): Dentin Caries

SUBMITTER: Yixin Zhang  

LAB HEAD: Dr Xian Peng

PROVIDER: PXD058535 | Pride | 2025-01-25

REPOSITORIES: Pride