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Creating Peptide Hydrazides via Intein Splicing for Native Chemical Ligation and Protein Labeling
Author(s) -
Santoleri Dominic A,
Liu Jun,
Ekanayake Oshini,
Rozovsky Sharon
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.602.6
Subject(s) - intein , protein tag , peptide , protein splicing , chemistry , hydrazide , rna splicing , combinatorial chemistry , biochemistry , trans splicing , native chemical ligation , ligation , computational biology , microbiology and biotechnology , biology , chemical synthesis , in vitro , gene , organic chemistry , recombinant dna , rna , fusion protein
Peptide hydrazides are powerful surrogates for the thioesters typically used in protein ligation as well as being an effective handle for labeling. Typical peptide hydrazide preparation relies on solid‐phase peptide synthesis, a technique limited by peptide size and solubility that also uses harsh chemicals. Here, a more effective and biochemically‐friendly means of generating these peptide hydrazides has been established involving split PhoRadA inteins, which only regain their splicing activity upon association with each other. This method also avoids several issues involved with full intein splicing, such as in vivo self‐cleavage and low expression levels. C‐Terminal peptide hydrazides are a powerful means of creating protein labels as we have shown them to react with small aldehyde‐containing molecules that can serve as a tag or a marker in analytical studies. Support or Funding Information Funding for this research project is provided through an NSF CAREER Award.