Peptide Cyclization Catalyzed by Cyanobactin Macrocyclases

Houssen, Wael E.

doi:10.1007/978-1-4939-9546-2_11

“…Peptide macrocyclization remains a challenge in chemistry [247] . In addition to YcaO mentioned in sections 5 (Figure 3C) and 17 (Figure 9D) for the modification of Cys, Ser, and Thr‐containing peptides, cyanobactin macrocyclases are promiscuous subtilisin‐like serine proteases that are present in the pathways of cyanobacterial peptides [248,249] . Examples include PatG, TruG, and PagG from the patellamide, [250,251] trunkamide, [252] and prenylagaramide [253] pathways, respectively.…”

Section: Cyclizationmentioning

confidence: 99%

“…[ 247 ] In addition to YcaO mentioned in section 5 ( Figure 3C ) and 17 ( Figure 9D ) for the modification of Cys, Ser, and Thr-containing peptides, cyanobactin macrocyclases are promiscuous subtilisin-like serine proteases that are present in the pathways of cyanobacterial peptides. [ 248 , 249 ] Examples include PatG, TruG, and PagG from the patellamide, [ 250 , 251 ] trunkamide, [ 252 ] and prenylagaramide [ 253 ] pathways, respectively. The PatG-related enzymes catalyze the N–C macrocyclization of short linear peptides composed of proteinogenic and non-proteinogenic amino acids, attached to a C-terminus recognition sequence that ranges from 3–7 amino acids and is different among G enzymes ( Figure 15A ).…”

Section: Cyclizationmentioning

confidence: 99%

Promiscuous Enzymes for Residue‐Specific Peptide and Protein Late‐Stage Functionalization

Alexander

¹

,

Elshahawi

²

2023

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The late‐stage functionalization of peptides and proteins holds significant promise for drug discovery and facilitates bioorthogonal chemistry. This selective functionalization leads to innovative advances in in vitro and in vivo biological research. However, it is a challenging endeavor to selectively target a certain amino acid or position in the presence of other residues containing reactive groups. Biocatalysis has emerged as a powerful tool for selective, efficient, and economical modifications of molecules. Enzymes that have the ability to modify multiple complex substrates or selectively install nonnative handles have wide applications. Herein, we highlight enzymes with broad substrate tolerance that have been demonstrated to modify a specific amino acid residue in simple or complex peptides and/or proteins at late‐stage. The different substrates accepted by these enzymes are mentioned together with the reported downstream bioorthogonal reactions that have benefited from the enzymatic selective modifications.

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“…13 Enzymes provide an additional tool to create libraries of cyclic peptides for drug discovery. Examples include subtiligase, [14][15] cyanobactin macrocyclases, 16 and cyclotide macrocyclases, [17][18][19][20] each of which has been exploited in drug discovery programs. As with chemical synthesis, each enzyme has its limits, including restricted substrate tolerance, reaction condition requirements, and others.…”

Section: Introductionmentioning

confidence: 99%

Expanding the Chemical Space of Synthetic Cyclic Peptides Using a Promiscuous Macrocyclase from Prenylagaramide Biosynthesis

Sarkar

¹

,

Gu

²

,

Schmidt

³

2020

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Cyclic peptides are excellent drug candidates, placing macrocyclization reactions at the apex of drug development. PatG and related dual-action proteases from cyanobactin biosynthesis are responsible for cleaving off the C-terminal recognition sequence and macrocyclizing the substrate to provide cyclic peptides. This reaction has found use in the enzymatic synthesis of diverse macrocycles. However, these enzymes function best on substrates that terminate with the non-proteinogenic thiazole/thiazoline residue, complicating synthetic strategies. Here, we biochemically characterize a new class of PatG-like macrocyclases that natively use proline, obviating the necessity of additional chemical or biochemical steps. We experimentally define the biochemical steps involved in synthesizing the widespread prenylagaramide-like natural products, including macrocyclization and prenylation. Using saturation mutagenesis, we show that macrocyclase PagG and prenyltransferase PagF are highly promiscuous, producing a library of more than 100 cyclic peptides and their prenylated derivatives in vitro. By comparing our results to known cyanobactin macrocyclase enzymes, we catalog a series of enzymes that collectively should synthesize most small macrocycles. Collectively, these data reveal that, by selecting the right cyanobactin macrocyclase, a large array of enzymatically synthesized macrocycles are accessible.

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“…13 Enzymes provide an additional tool to create libraries of cyclic peptides for drug discovery. Examples include subtiligase, [14][15] cyanobactin macrocyclases, 16 and cyclotide macrocyclases, [17][18][19][20] each of which has been exploited in drug discovery programs. As with chemical synthesis, each enzyme has its limits, including restricted substrate tolerance, reaction condition requirements, and others.…”

Section: Introductionmentioning

confidence: 99%

Expanding the chemical space of synthetic cyclic peptides using a promiscuous macrocyclase from prenylagaramide biosynthesis

Sarkar

¹

,

Gu

²

,

Schmidt

³

2020

Preprint

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Cyclic peptides are excellent drug candidates, placing macrocyclization reactions at the apex of drug development. PatG and related dual-action proteases from cyanobactin biosynthesis are responsible for cleaving off the C-terminal recognition sequence and macrocyclizing the substrate to provide cyclic peptides. This reaction has found use in the enzymatic synthesis of diverse macrocycles. However, these enzymes function best on substrates that terminate with the non-proteinogenic thiazole/thiazoline residue, complicating synthetic strategies. Here, we biochemically characterize a new class of PatG-like macrocyclases that natively use proline, obviating the necessity of additional chemical or biochemical steps. We experimentally define the biochemical steps involved in synthesizing the widespread prenylagaramide-like natural products, including macrocyclization and prenylation. Using saturation mutagenesis, we show that macrocyclase PagG and prenyltransferase PagF are highly promiscuous, producing a library of more than 100 cyclic peptides and their prenylated derivatives in vitro. By comparing our results to known cyanobactin macrocyclase enzymes, we catalog a series of enzymes that collectively should synthesize most small macrocycles. Collectively, these data reveal that, by selecting the right cyanobactin macrocyclase, a large array of enzymatically synthesized macrocycles are accessible.

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Peptide Cyclization Catalyzed by Cyanobactin Macrocyclases

Cited by 5 publications

References 46 publications

Promiscuous Enzymes for Residue‐Specific Peptide and Protein Late‐Stage Functionalization

Promiscuous Enzymes for Residue‐Specific Peptide and Protein Late‐Stage Functionalization

Expanding the Chemical Space of Synthetic Cyclic Peptides Using a Promiscuous Macrocyclase from Prenylagaramide Biosynthesis

Expanding the chemical space of synthetic cyclic peptides using a promiscuous macrocyclase from prenylagaramide biosynthesis

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