AI Article Synopsis
- - Sortase A (SrtA) is typically used for attaching labels to proteins but has limitations due to its reversibility and the need for a specific amino acid sequence (LPxTG).
- - Researchers discovered that SrtA can irreversibly link proteins/peptides with a thioester at one end to those with a Gly at the other end, allowing for flexibility beyond the traditional LPxTG requirement.
- - This new approach allows for the easy creation of proteins with various labels, including modified proteins like phosphorylated and methylated histones, demonstrating that substrate modification can improve existing enzymatic techniques.
Article Abstract
Sortase A (SrtA)-mediated ligation, a popular method for protein labeling and semi-synthesis, is limited by its reversibility and dependence on the LPxTG motif, where "x" is any amino acid. Here, we report that SrtA can mediate the efficient and irreversible ligation of a protein/peptide containing a C-terminal thioester with another protein/peptide bearing an N-terminal Gly, with broad tolerance for a wide variety of LPxT-derived sequences. This strategy, the thioester-assisted SrtA-mediated ligation, enabled the expedient preparation of proteins bearing various N- or C-terminal labels, including post-translationally modified proteins such as the Ser139-phosphorylated histone H2AX and Lys9-methylated histone H3, with less dependence on the LPxTG motif. Our study validates the chemical modification of substrates as an effective means of augmenting the synthetic capability of existing enzymatic methods.
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| http://dx.doi.org/10.1002/anie.202201887 | DOI Listing |
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Article Synopsis
- - Sortase A (SrtA) is typically used for attaching labels to proteins but has limitations due to its reversibility and the need for a specific amino acid sequence (LPxTG).
- - Researchers discovered that SrtA can irreversibly link proteins/peptides with a thioester at one end to those with a Gly at the other end, allowing for flexibility beyond the traditional LPxTG requirement.
- - This new approach allows for the easy creation of proteins with various labels, including modified proteins like phosphorylated and methylated histones, demonstrating that substrate modification can improve existing enzymatic techniques.
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