XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
82.
CRH TraNSGLYCOSYLASES CATALYZE INTER POLYMERIC LINKAGES IN
FUNGAL CELL WALL
Marián Mazáň 1 , Noelia Blanco 2 , Javier Arroyo 2 and Vladimír Farkaš 1
1
Institute of Chemistry - Center for Glycomics, Slovak Academy of Sciences,
Dúbravská cesta 9, 84538 Bratislava, Slovakia
2
Departamento de Microbiología II, Facultad de Farmacia,
Universidad Complutense de Madrid, 28040 Madrid, Spain
The cell wall of Saccharomyces cerevisiae is an elastic structure that provides osmotic
and physical protection and determines the shape of the cell. Crh1 and Crh2 are GPIlinked
cell wall proteins required for the formation of linkages between chitin and β-1,3-
glucose branches of β-1,6-glucan in the cell wall of budding yeast. According to the CAZY
database, they belong to the glycoside hydrolase family 16.
We developed a fluorescent in vitro assay system for determination of the transglycosylating
activity of Crh proteins. His-tagged Crh1p and Crh2p of S. cerevisiae were heterologously
expressed in Pichia pastoris, purified on Ni-column and their basic biochemical
characteristics were investigated.
The following properties of Crh proteins were determined: pH and temperature optima,
donor and acceptor substrate specificity, effectors, thermal stability and mode of action.
Acknowledgement: This work was supported by grant no. 2/0011/09 from the Slovak
Grant Agency for Science VEGA.
XXII. Biochemistry Congress, Martin
205