Cellular Localization and Biochemical Characterization of a Chimeric Fluorescent Protein Fusion of Arabidopsis Cellulose Synthase-Like A2 Inserted into Golgi Membrane

Table 1

Mannan synthase activity in wild-type and AtCslA2-GFP transformed tobacco protoplasts and effect of protease K with or without Triton X-100. The reaction mixture contained the following: 0.20 nmol of GDP-D-[U-¹⁴C]mannose, 20 nmol GDP-D-mannose, and 10 μL (130 μg of protein) of total membrane fraction isolated from wild-type and AtCslA2-GFP transformed tobacco protoplasts, in 0.1 M sodium-phosphate buffer, pH 7.2, in a total volume of 50 μL. Reaction time at 27°C was 30 min. Data are the means ± standard deviation of three independent replicates (𝑛=3).

Protoplasts	Bq	Specific activity (nmol·min/mg of protein)
Wild-type	3.9±0.2	370±25
AtCslA2-GFP transformed	5.4±0.3	510±32
+ Proteinase K	4.6±0.3	438±30
+ Triton X-100	2.2±0.1	209±16
+ Proteinase K + Triton X-100	0.8±0.1	76±5