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Ligation of Soluble but Unreactive Peptide Segments in the Chemical Synthesis of Haemophilus Influenzae DNA Ligase
Author(s) -
Zhang Baochang,
Deng Qiang,
Zuo Chong,
Yan Bingjia,
Zuo Chao,
Cao XiuXiu,
Zhu Ting F.,
Zheng JiShen,
Liu Lei
Publication year - 2019
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201905149
Subject(s) - chemistry , chemical ligation , peptide , native chemical ligation , dna ligase , ligation , guanidine , combinatorial chemistry , solubility , ligase chain reaction , dna , peptide synthesis , biophysics , biochemistry , chemical synthesis , organic chemistry , biology , microbiology and biotechnology , in vitro , polymerase chain reaction , multiplex polymerase chain reaction , gene
During the total chemical synthesis of the water‐soluble globular Haemophilus Influenzae DNA ligase (Hin‐Lig), we observed the surprising phenomenon of a soluble peptide segment that failed to undergo native chemical ligation. Based on dynamic light scattering and transmission electron microscopy experiments, we determined that the peptide formed soluble colloidal particles in a homogeneous solution containing 6 m guanidine hydrochloride. Conventional peptide performance‐improving strategies, such as installation of a terminal/side‐chain Arg tag or O ‐acyl isopeptide, failed to enable the reaction, presumably because of their inability to disrupt the formation of soluble colloidal particles. However, a removable backbone modification strategy recently developed for the synthesis of membrane proteins did disrupt the formation of the colloids, and the desired ligation of this soluble but unreactive system was eventually accomplished. This work demonstrates that an appropriate solution dispersion state, in addition to good peptide solubility, is a prerequisite for successful peptide ligation.