Premium
15 N labeling method of peptides using a thioredoxin gene fusion expression system: an application to ACTH‐(1–24)
Author(s) -
Uegaki Koichi,
Nemoto Nobuaki,
Shimizu Masato,
Wada Takashi,
Kyogoku Yoshimasa,
Kobayashi Yuji
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01459-4
Subject(s) - thioredoxin , enteropeptidase , peptide , chemistry , fusion protein , gene expression , biochemistry , fusion gene , microbiology and biotechnology , gene , adrenocorticotropic hormone , biology , hormone , recombinant dna
For structure analysis of peptides by multinuclear NMR, stable isotope‐labeled samples are required. A direct over‐expression system by E. coli cells does not work for that purpose because of rapid degradation of the peptides and/or the mRNA in host cells. We here developed an over‐expression system by means of thioredoxin gene fusion system. The fused protein composed of thioredoxin and the objective peptide was expressed in E. coli and then the peptide part was released by enterokinase. This system was successfully applied for the production of 15 N‐labeled human adrenocorticotropic hormone fragment (ACTH‐(1‐24)) as needed for multinuclear NMR analysis.