Premium
Efficient Generation of Hydrazides in Proteins by RadA Split Intein
Author(s) -
Liu Jun,
Ekanayake Oshini,
Santoleri Dominic,
Walker Kelsi,
Rozovsky Sharon
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900160
Subject(s) - intein , chemistry , cleavage (geology) , native chemical ligation , bioconjugation , combinatorial chemistry , palmitoylation , biochemistry , in vitro , cysteine , chemical synthesis , biology , enzyme , rna , paleontology , fracture (geology) , rna splicing , gene
Abstract Protein C‐terminal hydrazides are useful for bioconjugation and construction of proteins from multiple fragments through native chemical ligation. To generate C‐terminal hydrazides in proteins, an efficient intein‐based preparation method has been developed by using thiols and hydrazine to accelerate the formation of the transient thioester intermediate and subsequent hydrazinolysis. This approach not only increases the yield, but also improves biocompatibility. The scope of the method has been expanded by employing Pyrococcus horikoshii RadA split intein, which can accommodate a broad range of extein residues before the site of cleavage. The use of split RadA minimizes premature intein N cleavage in vivo and offers control over the initiation of the intein N cleavage reaction. It is expected that this versatile preparation method will expand the utilization of protein C‐terminal hydrazides in protein preparation and modification.