Convergent Versus Sequential Protein Synthesis: The Case of Ubiquitinated and Glycosylated H2B

Abstract

The chemical synthesis of a protein from four fragments or more applying native chemical ligation could be achieved stepwise, in one-pot, convergently, or on a solid support. With the increasing demands of applying protein synthesis to highly complex targets, examining these approaches becomes essential to achieve highly efficient synthesis. Different chemical synthetic strategies are compared for the preparation of the H2B protein having different post-translational modifications. The analogues include H2B that is ubiquitinated at Lys34, Lys120, glycosylated at Ser112, and doubly modified with ubiquitin and N-acetylglucosamine. This study demonstrates that the applied convergent strategy for the synthesis of most of these complex targets was better than the one-pot approach in terms of yield and purity. Some guidelines are offered for future synthetic endeavors of similar challenging proteins.

Aligator: A computational tool for optimizing total chemical synthesis of large proteins

“…While one-pot and solid-phase ligation strategies can be used, convergent synthesis is generally the most efficient approach 91 . Consideration of the optimal ligation order will depend on the specific terminal protecting groups used, as well as the arrangement of residues requiring desulfurization.…”

Chemistry‐Driven Epigenetic Investigation of Histone and DNA Modifications

“…The results revealed that the ubiquitin protruded between the DNA chains. K120‐ubiquitinated H2B was chemically synthesized by these two groups ,. Liu's group developed a new activation method, in which peptide−Dbz linker was converted to peptide benzotriazole by sodium nitrite after cleavage from the resin .…”

Chemical Synthesis of Diubiquitin‐Based Photoaffinity Probes for Selectively Profiling Ubiquitin‐Binding Proteins

“…Because the binding between Ub and UBDs usually involves the Ile 44 hydrophobic patch of Ub, [12] we put ap hoto-crosslinking phenyl azide group at the Ala 46 that neighbors Ile 44. [14] For example,toprepare 3,wedivided it into three segments: 4, 5, and 6 ( Figure 2A). [13] As ar esult, we carried out protein total synthesis, which allows exquisite control at the atomic level.…”