Chemoselective Ligation of Peptide Phenyl Esters with N‐Terminal Cysteines

Fang, Ge‐Min; Cui, Hongyang; Zheng, Ji‐Min; Liu, Lei

doi:10.1002/cbic.201000165

Cited by 32 publications

(21 citation statements)

References 49 publications

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“…Fortunately, Liu and coworkers demonstrated, with imidazole activation, a C- terminal phenyl ester could successfully participate in an NCL in the place of a C -terminal thioester. 18 …”

Section: Resultsmentioning

confidence: 99%

A Fully Synthetic and Biochemically Validated Phosphatidyl Inositol-3-Phosphate Hapten via Asymmetric Synthesis and Native Chemical Ligation

et al. 2013

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We report the synthesis and biochemical validation of a phosphatidyl inositol-3 phosphate (PI3P) immunogen. The inositol stereochemistry was secured through peptide-catalyzed asymmetric phosphorylation catalysis, and the subsequent incorporation of a cysteine residue was achieved by native chemical ligation (NCL). Conjugation of the PI3P hapten to maleimide activated keyhole limpet hemocyanin (KLH) provided a PI3P immunogen, which was successfully used to generate selective PI3P antibodies. The incorporation of a sulfhydryl nucleophile into a phosphoinositide hapten demonstrates a general strategy to reliably access phosphoinositide immunogens.

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“…Fortunately, Liu and coworkers demonstrated, with imidazole activation, a C- terminal phenyl ester could successfully participate in an NCL in the place of a C -terminal thioester. 18 …”

Section: Resultsmentioning

confidence: 99%

A Fully Synthetic and Biochemically Validated Phosphatidyl Inositol-3-Phosphate Hapten via Asymmetric Synthesis and Native Chemical Ligation

et al. 2013

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“…Many types of activating groups have been used for peptide ligations, including thioesters [85], acyl azides [86][87][88][89][90], acid chlorides [91], acyl isoureas [92], acyl imidazoles [93], and aromatic esters [94]. Of all the possibilities, however, peptide thioesters have become the most widely used (Fig.…”

Section: New Methods For Activated Peptide Synthesismentioning

confidence: 99%

New Methods for Chemical Protein Synthesis

Guan

Chaffey

Zeng

2014

Topics in Current Chemistry

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Chemical protein synthesis is a useful tool to generate pure proteins which are otherwise difficult to obtain in sufficient amounts for structure and property analysis. Additionally, because of the precise and flexible nature of chemical synthesis, it allows for controllable variation of protein sequences, which is valuable for understanding the relationships between protein structure and function. Despite the usefulness of chemical protein synthesis, it has not been widely adopted as a tool for protein characterization, mainly because of the lack of general and efficient methods for the preparation and coupling of peptide fragments and for the folding of polypeptide chains. To address these issues, many new methods have recently been developed in the areas of solid-phase peptide synthesis, peptide fragment assembly, and protein folding. Here we review these recent technological advances and highlight the gaps needing to be addressed in future research.

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“…Synthesis of the N-terminal peptide segment (segment 2) followed the standard Fmoc strategy [22,23]. Wang resin was initially swelled with DCM/DMF (1/1) for 3 h. To pre-activate the first protected amino acid, 3.6 equiv HBTU, 4 equiv HOBt, 8 equiv DIEA and 0.1 equiv DMAP were added to a solution of 4 equiv protected amino acid (0.4 M) in DMF.…”

Section: Synthesis Of Avc*gqrsggidvktksklvc-nh 2 (Segment 2) Followinmentioning

confidence: 99%

Hexafluoro-2-propanol as a potent cosolvent for chemical ligation of membrane proteins

2011

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The study on membrane proteins is an important challenge mainly because of their very poor solubility in various solvents. The traditional recombinant expression strategy and the native chemical ligation method both have difficulty in generating sufficient amounts of desired proteins with high efficiency. Previous studies have shown that multiply fluorinated alcohols exhibit good ability to dissolve difficult peptide sequences, especially hexafluoro-2-propanol (HFIP). In the present study we systematically studied the capability of solvents containing different percentage of HFIP in dissolving transmembrane peptides. Through both HPLC and UV analyses we concluded that 60% HFIP/8 M urea constituted a good solvent system. In this solvent system we also optimized conditions to perform native chemical ligation (NCL). Under the optimized conditions we successfully achieved NCL's for both dipeptide formation and the synthesis of a model protein (Trifolitoxin). These results suggested that HFIP was a potential cosolvent that could be used in the ligation of poorly soluble peptides for the generation of membrane proteins. membrane protein, native chemical ligation, alternative solvent, hexafluoro-2-propanol

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Chemoselective Ligation of Peptide Phenyl Esters with N‐Terminal Cysteines

Cited by 32 publications

References 49 publications

A Fully Synthetic and Biochemically Validated Phosphatidyl Inositol-3-Phosphate Hapten via Asymmetric Synthesis and Native Chemical Ligation

A Fully Synthetic and Biochemically Validated Phosphatidyl Inositol-3-Phosphate Hapten via Asymmetric Synthesis and Native Chemical Ligation

New Methods for Chemical Protein Synthesis

Hexafluoro-2-propanol as a potent cosolvent for chemical ligation of membrane proteins

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