| RRC ID |
4956
|
| 著者 |
Ebhardt HA, Xu Z, Fung AW, Fahlman RP.
|
| タイトル |
Quantification of the post-translational addition of amino acids to proteins by MALDI-TOF mass spectrometry.
|
| ジャーナル |
Anal Chem
|
| Abstract |
Aminoacyl-tRNA protein transferases catalyze the post-translational addition of amino acids to proteins. The eubacterial leucyl/phenylalanyl-tRNA-protein transferase (L/F transferase) catalyzes the transfer of leucine or phenylalanine from their respective aminoacylated tRNAs to the N-termini of substrate proteins possessing an N-terminal lysine or arginine amino acid. Conventional assays to quantify L/F transferase activity involve measuring radioactive amino acid incorporation into substrate proteins. We have developed a quantitative matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry procedure to measure the enzymatic activity of L/F transferase. The procedure utilizes stable isotope labeled substrate and internal standard peptides. The method is used to determine the kinetic parameters of k(cat) and K(m) for the enzymatic transfer of phenylalanine and three unnatural amino acid derivatives from an aminoacyl-tRNA to a peptide substrate.
|
| 巻・号 |
81(5)
|
| ページ |
1937-43
|
| 公開日 |
2009-3-1
|
| DOI |
10.1021/ac802423d
|
| PII |
10.1021/ac802423d
|
| PMID |
19186990
|
| MeSH |
Amino Acids / chemistry*
Protein Processing, Post-Translational*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Transferases / metabolism
|
| IF |
6.785
|
| 引用数 |
10
|
|
WOS 分野
|
CHEMISTRY, ANALYTICAL
|
| リソース情報 |
| 原核生物(大腸菌) |
ASKA(-) collection? |
