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. 1997 Nov 3;139(3):625–638. doi: 10.1083/jcb.139.3.625

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7B2-induced activation of proPC2 from Golgi membranes. (a) Subcellular fractions (3 μg protein) corresponding to the supernatant of the first centrifugation (S1) and to Golgi- and ER-enriched membranes were incubated at pH 5 at 37°C for 30 min in the presence or absence of 200 nM recombinant 21-kD 7B2 (21 kD) and of 1 μM CT peptide (CT). The reaction was stopped by the addition of Laemmli sample buffer and boiled for 5 min before PC2 immunoblotting. (b) Subcellular fractions (1 μg protein) were incubated under the same conditions as in a and in the presence of 200 μM Pyr-Glu-Arg-Thr-Lys-Arg-AMC. Fluorescence was measured and PC2 specific activity was calculated. Black bars correspond to the control incubations, grey bars to the incubations with the recombinant 21-kD 7B2, and white bars to the incubation with the recombinant 21-kD 7B2 and the CT peptide.

7B2-induced activation of proPC2 from Golgi membranes. (a) Subcellular fractions (3 μg protein) corresponding to the supernatant of the first centrifugation (S1) and to Golgi- and ER-enriched membranes were incubated at pH 5 at 37°C for 30 min in the presence or absence of 200 nM recombinant 21-kD 7B2 (21 kD) and of 1 μM CT peptide (CT). The reaction was stopped by the addition of Laemmli sample buffer and boiled for 5 min before PC2 immunoblotting. (b) Subcellular fractions (1 μg protein) were incubated under the same conditions as in a and in the presence of 200 μM Pyr-Glu-Arg-Thr-Lys-Arg-AMC. Fluorescence was measured and PC2 specific activity was calculated. Black bars correspond to the control incubations, grey bars to the incubations with the recombinant 21-kD 7B2, and white bars to the incubation with the recombinant 21-kD 7B2 and the CT peptide.