Synthesis of Post-translational Modifier Protein NEDD8 via Ligation of Peptide Hydrazides

Abstract

As an important ubiquitin-like modifier protein in eukaryotic organisms, NEDD8 (neural precursor cell expressed developmentally down-regulated 8) is involved in regulating a series of important life processes in cells. Nowadays, the major method for obtaining NEDD8 is recombinant protein expression. However, the yield is relatively low and the recombined tag for purification needs to be removed in an extra step. In present work, NEDD8 protein was first synthesized in homogeneity by using high temperature assisted solid-phase peptide synthesis (SPPS) combining with the one-pot ligation-desulfurization strategy. This method lays the foundation for the study of NEDD8 modified proteins in future.

DIC/Oxyma‐based accelerated synthesis and oxidative folding studies of centipede toxin RhTx

“…However, there are potential explosion risks during SPPS using the HOBt and HOBt‐based coupling reagents, especially in large‐scale synthesis 19,21 . Recently, Oxyma [ethyl 2‐cyano‐2‐(hydroxyimino)acetate] was developed as the relatively safe racemic inhibitor, which has been used in the carbodiimide‐based synthesis of ubiquitin protein segments, polyubiquitin chains, liraglutide, and so forth 21–26 …”

“…19,21 Recently, Oxyma [ethyl 2-cyano-2-(hydroxyimino)acetate] was developed as the relatively safe racemic inhibitor, which has been used in the carbodiimide-based synthesis of ubiquitin protein segments, polyubiquitin chains, liraglutide, and so forth. [21][22][23][24][25][26] Compared with traditional racemic inhibitors such as HOBt, Oxyma has the higher coupling efficiency, lower risk of explosion, and lower rate of racemization during peptide coupling reaction. 21 The N,N 0 -diisopropylcarbodiimide (DIC)/Oxyma coupling system has been demonstrated to be compatible with microwave-assisted SPPS and reduce racemization under high temperature (from 70 C to 90 C).…”