1. Preparation ofN-Acylated Proteins Modified with Fatty Acids Having a Specific Chain Length Using an Insect Cell-Free Protein Synthesis System
- Author
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Toshihiko Utsumi, Toru Ezure, Takashi Suzuki, Masamitsu Shikata, Eiji Ando, Susumu Tsunasawa, Masaaki Ito, and Osamu Nishimura
- Subjects
Stereochemistry ,Molecular Sequence Data ,Spodoptera ,Biology ,Applied Microbiology and Biotechnology ,Biochemistry ,Cell Line ,Analytical Chemistry ,Acylation ,Protein biosynthesis ,Animals ,Amino Acid Sequence ,Molecular Biology ,chemistry.chemical_classification ,Cell-Free System ,Fatty Acids ,Organic Chemistry ,Proteins ,Fatty acid ,Acetylation ,General Medicine ,Protein engineering ,Chain length ,chemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Saturated fatty acid ,Free fatty acid receptor ,lipids (amino acids, peptides, and proteins) ,Biotechnology ,Polyunsaturated fatty acid - 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.
- Published
- 2007
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