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14C]cysteine (303 mCi/mmol, NEN DuPont; reduced by DTT)]} or proline (103 Ci/mmo, Amersham). Samples taken at various time points were spotted on Whatman 3MM filter disks, presoaked in 10% trichloroacetic acid. The disks were washed and radioactivity was measured by liquid scintillation counting.
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6-ProRS was judged to be at least 99% pure by Coomassie Blue staining after SDS-polyacrylamide gel electrophoresis (PAGE).
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6-ProRS as described (7).
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note
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We thank C. A. Hutchison III for sharing unpublished results, and A. Ambrogelly, C. Becker, G. Raczniak, and D. Tumbula for assistance and advice. H.D.B. is a European Molecular Biology Organization postdoctoral fellow, M.I. is supported by an Investigator Fellowship from the Alfred Benzon Foundation and D.S. by grants from the National Institute of General Medical Sciences, NASA, and the Department of Energy.
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