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Detection of DNA Damage in Cultured Human Fibroblasts Induced by Methyl Linoleate Hydroperoxide
Authors:Tsutomu Nakayama  Motohisa Kaneko  Masahiko Kodama
Affiliation:Biophysics Division, National Cancer Center Research Institute, Tsukiji 5–1–1, Chuo-ku, Tokyo 104, Japan
Abstract:To establish a strategy to generate N-acylated proteins modified with fatty acids having a specific chain length, tGelsolin-streptag, an epitope-tagged model protein having an N-myristoylation motif, was synthesized using an insect cell-free protein synthesis system in the presence of acyl-CoA with various fatty acid chain lengths. It was found that the fatty acid species attached to the N-termini fully depended on the acyl-CoA species added to the reaction mixture. N-Acylated proteins with fatty acid chain lengths of 8, 10, 12, and 14 were generated successfully.
Keywords:N-acylation  insect cell-free protein synthesis system  protein engineering  saturated fatty acid
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