Preparation of N-acylated proteins modified with fatty acids having a specific chain length using an insect cell-free protein synthesis system.
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.
Suzuki
Takashi
T
Life Science Laboratory, Analytical and Measuring Instruments Division, Shimadzu Corporation, Japan. t-suzuki@shimadzu.co.jp
Ito
Masaaki
M
Ezure
Toru
T
Shikata
Masamitsu
M
Ando
Eiji
E
Utsumi
Toshihiko
T
Tsunasawa
Susumu
S
Nishimura
Osamu
O
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018411:Spodoptera
D005227:Fatty Acids; D011506:Proteins