Concentrations of various additions to lysis buffer for optimal C. neoformans (1,3)β-glucan synthase activitya

AdditionConcn testedOptimal concn
MgCl2 (mM)1-105
CaCl2 (mM)1-107
NaF (mM)1-52
Phosphatase inhibitor (% [vol/vol])0.01-10.1
Glycerol (% [vol/vol])10-5020
Sucrose (M)0.1-1.00.5
PMSF (mM)0.05-10.1
DTT (mM)1-1001
Protease inhibitor (% [vol/vol])0.01-10.1
BSA (mg/ml)1-5
EDTA (mM)1-10050
EGTA (mM)1-10020
GTPγS (μM)5-20060
  • a H99 cells were lysed as described in Materials and Methods, and the optimal concentration of each addition to the lysis buffer resulting in maximal activity was determined as described in Materials and Methods and the text.