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. 2000 Oct 16;151(2):289–296. doi: 10.1083/jcb.151.2.289

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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[³H]GDP release from Ypt1p in the presence of cell lysates prepared from ts mutants that are defective in different TRAPP subunits. His₆-Ypt1p preloaded with [³H]GDP was incubated with cell lysates for 10 min at 30°C (A) or 37°C (B) as described in the Materials and Methods. Lysates assayed at 37°C were pre-incubated for 5 min at 37°C immediately before the reaction was performed. The average intrinsic rate of [³H]GDP release from Ypt1p was measured in the presence of BSA (4 mg/ml) and the value obtained (15.5 ± 2% at 30°C; 28.0 ± 2% at 37°C) was subtracted as background. The numbers reported are the average of two separate experiments.

[³H]GDP release from Ypt1p in the presence of cell lysates prepared from ts mutants that are defective in different TRAPP subunits. His₆-Ypt1p preloaded with [³H]GDP was incubated with cell lysates for 10 min at 30°C (A) or 37°C (B) as described in the Materials and Methods. Lysates assayed at 37°C were pre-incubated for 5 min at 37°C immediately before the reaction was performed. The average intrinsic rate of [³H]GDP release from Ypt1p was measured in the presence of BSA (4 mg/ml) and the value obtained (15.5 ± 2% at 30°C; 28.0 ± 2% at 37°C) was subtracted as background. The numbers reported are the average of two separate experiments.