Recent applications of enzymatic peptide synthesis

Bongers, Jacob; Heimer, Edgar P.

doi:10.1016/0196-9781(94)90189-9

Cited by 83 publications

(38 citation statements)

References 85 publications

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“…The second 18 O atom-incorporation reaction (reaction 2) is essentially the reverse reaction of peptide-bond hydrolysis or the peptide-bond formation reaction (protease catalysts accelerate the forward and reverse reactions). There are in fact numerous experimental studies that verify the occurrence of the proteasecatalyzed peptide-bond formation reaction (for example, Bongers & Heimer, 1994). The only difference between reaction 2 and the peptide-bond formation reaction is the reacting reagent and the nucleophilic atom on that reagent.…”

Section: Enzymology Of Proteolytic 18 O-labeling Reactionmentioning

confidence: 97%

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics

Miyagi

Rao

2006

Mass Spectrometry Reviews

168

208

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A number of proteomic techniques have been developed to quantify proteins in biological systems. This review focuses on the quantitative proteomic technique known as "proteolytic 18O-labeling." This technique utilizes a protease and H(2)18O to produce labeled peptides, with subsequent chromatographic and mass spectrometric analysis to identify and quantify (relative) the proteins from which the peptides originated. The technique determines the ratio of individual protein's expression level between two samples relative to each other, and can be used to quantitatively examine protein expression (comparative proteomics) and post-translational modifications, and to study protein-protein interactions. The present review discusses various aspects of the 18O-labeling technique, including: its history, the advantages and disadvantages of the proteolytic 18O-labeling technique compared to other techniques, enzymatic considerations, the problem of variable incorporation of 18O atoms into peptides with a discussion on recent advancements of the technique to overcome it, computational tools to interpret the data, and a review of the biological applications.

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Section: Enzymology Of Proteolytic 18 O-labeling Reactionmentioning

confidence: 97%

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics

Miyagi

Rao

2006

Mass Spectrometry Reviews

168

208

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“…256 Trypsin and chymotrypsin have also been used to catalyze incorporation of sterically demanding α,α-disubstituted-fluoroalkyl amino acids into peptides using reactive 4-guanidinophenyl esters as acyl donors. 264 The use of proteases for peptide synthesis has been extensively reviewed, [265][266][267] with solid-phase approaches having been used to prepare peptide targets that are difficult to synthesize in solution phase due to competing hydrolytic pathways. 268 These solidphase systems have the advantage that a large excess of acyl donors can be used to drive peptide bond-forming reactions to completion, with the solid support serving to suppress ionization of the free amino group, and improve solvation of hydrophilic acyl donors.…”

Section: Amide Bond-forming Reactions For the Synthesis Of Peptides Amentioning

confidence: 99%

6.11 N-Acylation Reactions of Amines

Taylor

Bull

2014

Comprehensive Organic Synthesis II

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“…[6] Since peptide bond synthesis is thermodynamically unfavourable in the presence of water, the use of neat organic solvents with very low water activity can be beneficial for peptide coupling. For instance, the industrial serine protease Alcalase has been applied for peptide synthesis under nearly anhydrous conditions.…”

Section: Introductionmentioning

confidence: 99%

Peptiligase, an Enzyme for Efficient Chemoenzymatic Peptide Synthesis and Cyclization in Water

Toplak

Nuijens²,

Quaedflieg³

et al. 2016

Adv Synth Catal

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We describe a novel, organic cosolventstable and cation-independent engineered enzyme for peptide coupling reactions. The enzyme is a variant of a stable calcium-independent mutant of subtilisin BPN', with the catalytic Ser212 mutated to Cys and Pro216 converted to Ala. The enzyme, called peptiligase, catalyzes exceptionally efficient peptide coupling in water with a surprisingly high synthesis over hydrolysis (S/H) ratio. The S/H ratio of the peptide ligation reaction is correlated to the length of the peptide substrate and proved to be > 100 for the synthesis of a 13-mer peptide, which corresponds to > 99% conversion to the ligated peptide product and < 1% hydrolytic side-reaction. Furthermore, peptiligase does not require a particular recognition motif resulting in a broadly applicable and traceless peptide ligation technology. Peptiligase is very robust, easy to produce in Bacillus subtilis, and its purification is straightforward. It shows good activity and stability in the presence of organic cosolvents and chelating or denaturing agents, enabling the ligation of poorly soluble (hydrophobic) or folded peptides. This enzyme could be useful for the (industrial) synthesis of diverse (pharmaceutical) peptides. In addition, peptiligase is able to efficiently catalyze headto-tail peptide cyclization reactions.

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Recent applications of enzymatic peptide synthesis

Cited by 83 publications

References 85 publications

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics

6.11 N-Acylation Reactions of Amines

Peptiligase, an Enzyme for Efficient Chemoenzymatic Peptide Synthesis and Cyclization in Water

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Recent applications of enzymatic peptide synthesis

Cited by 83 publications

References 85 publications

Proteolytic 18O‐labeling strategies for quantitative proteomics

Proteolytic 18O‐labeling strategies for quantitative proteomics

6.11 N-Acylation Reactions of Amines

Peptiligase, an Enzyme for Efficient Chemoenzymatic Peptide Synthesis and Cyclization in Water

Contact Info

Product

Resources

About

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics

Proteolytic ¹⁸O‐labeling strategies for quantitative proteomics