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Figure 6


Fig. 6. Plasma membrane levels of Chs3p and biological activity. (A) Quantification of Chs3p-3xHA levels in the plasma membrane fraction after discontinuous sucrose gradients and western analysis as in Fig. 2C. Protein extracts were obtained from the JAY29 (chs4{Delta}) strain transformed with the indicated plasmids. Data represent the amount of Chs3p-3xHA localized in fractions 5-7 corresponding to the plasma membrane and are the average of three experiments. They are compared with resistance to 0.025 mg/ml Calcofluor and chitin levels (nmoles/100 mg cells), measured as indicated in the Materials and Methods. Calcofluor staining of these strains is shown in the lower images. (B) Distribution of Chs3p-3xHA in discontinuous sucrose gradients using extracts obtained from the chs4{Delta} and pRS424-CHS4 strains as in A. Lower panel represents specific CSIII activity (mU) in each fraction.





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