Ligation methods

Elegant work in which the chemical ligation methodology was used for the preparation of a polymer-modified protein was reported by Kochendoerfer 

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Fig. 4. Lysine-based core peptide 5 coupled with the model peptide VA 20 (represented by open boxes) by various ligation methods [17]...

Synthesis of Lipidated Proteins Using Ligation Methods 568... 

Scheme 33 Expressed protein ligation (EPL) as an example for a protein ligation method generating a native bond (Left). MIC ligation generating a non-native bond between the two coupied segments (right).

Shogren-Rnaak, M.A. and Peterson, C.L. (2004) Creating designer histones hy native chemical ligation. Methods in Enzymohgy, 375, 62—76. 

Chemical ligation methods for peptide synthesis using thioester chemistry in solution have been previously documented (see Vol. E 22a, Section 4.1.5). Generalized procedures for solid-phase ligation have been developed that simplify the overall procedure. One method uses a safety-catch acid labile linker at the C-terminus and was used for the synthesis of a 71-amino acid chemokine, vMIP I (Section 5.3.2.1). Another procedure uses a selectively cleavable glycolate ester linkage (Section 5.3.2.2). 

In addition to homodetic cyclic peptides, chemical ligation methods have been developed that lead to heterodetic cyclic peptides containing thiazolidine. 158-161 ... 

Scheme 16 Chemoselective Ligation Methods for Protein Designl18-24 951...

Another versatile chemoselective ligation method for the efficient synthesis of TASP[98] molecules is based on the rapid and selective reaction of thiols with activated double bonds, e.g. in a maleimide group (Scheme 17).[106-108] For this purpose, the cyclic decameric template is functionalized with four maleimide residues, which allow reaction with thiol-containing peptide fragments in aqueous solution. 

A plethora of ligation methods and many variations have been developed.1"-1121 Many nucleophilic or electrophilic agents with different spacers are currently commercially available as cross-linking reagents for protein conjugation. Furthermore, placement of the nucleophile-electrophile pair on the core and many possible positions on the peptide create... 

Blaschke, U. K., Silberstein, J., and Muir, T. W. (2000) Protein engineering by expressed protein ligation. Methods Enzymol. 328, 478-496. 

Protocol 4 Splinted RNA ligation method for producing FRET-labeled telomerase RNA... 

Moore, M., and Query, C. C. (2000). Joining of RNAs by splinted ligation. Methods... 

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