A Systematic Study of Selective Protein Glycation

Sjoblom, Nicole M.; Kelsey, Maxfield M G; Scheck, Rebecca A.

doi:10.1002/ange.201810037

“…Our previous work has revealed that primary sequence, on its own, shapes the distribution of resulting AGEs 29 . In our prior study, AGE products with an [M + 54] mass change, associated with formation of the MGH isomers, were the most frequently observed 29 .…”

Section: Resultsmentioning

confidence: 99%

“…Our previous work has revealed that primary sequence, on its own, shapes the distribution of resulting AGEs 29 . In our prior study, AGE products with an [M + 54] mass change, associated with formation of the MGH isomers, were the most frequently observed 29 . Of these, MGH-1 is known to be the most thermodynamically stable and has also been frequently detected in biological studies of glycation 8 , 14 , 34 , 38 , 39 .…”

Section: Resultsmentioning

confidence: 99%

“…1 and 2 ). Building on this observation, we designed a combinatorial library based on an Arg-containing sequence from human serum albumin (HSA) that we 29 , and others 25 , have found to be glycated in vitro (see Methods). After MGO treatment, sequences that produced MGH-1 were identified using a primary antibody specific for MGH-1 and subsequent colorimetric detection (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…2 , Supplementary Figs. 7 and 8 , Supplementary Table 2 ) 29 .…”

Section: Resultsmentioning

confidence: 99%

“…Although past work has cataloged numerous instances of preferential glycation 20 – 28 , there has been little investigation of the mechanism through which specific AGEs form on proteins 25 . Additionally, it remains an open question as to what controls the overall propensity for certain proteins to become glycated at certain sites 29 , 30 . Recent work from our lab has demonstrated that protein sequence is a primary determinant that governs the likelihood that a site will become glycated, and has also revealed that both sequence and folded structure sculpt the distinct complement of AGEs that form 29 .…”

Section: Introductionmentioning

confidence: 99%

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Synergistic sequence contributions bias glycation outcomes

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The methylglyoxal-derived hydroimidazolone isomer, MGH-1, is an abundant advanced glycation end-product (AGE) associated with disease and age-related disorders. As AGE formation occurs spontaneously and without an enzyme, it remains unknown why certain sites on distinct proteins become modified with specific AGEs. Here, we use a combinatorial peptide library to determine the chemical features that favor MGH-1. When properly positioned, tyrosine is found to play an active mechanistic role that facilitates MGH-1 formation. This work offers mechanistic insight connecting multiple AGEs, including MGH-1 and carboxyethylarginine (CEA), and reconciles the role of negative charge in influencing glycation outcomes. Further, this study provides clear evidence that glycation outcomes can be influenced through long- or medium-range cooperative interactions. This work demonstrates that these chemical features also predictably template selective glycation on full-length protein targets expressed in mammalian cells. This information is vital for developing methods that control glycation in living cells and will enable the study of glycation as a functional post-translational modification.

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Visualization of Protein‐Specific Glycation in Living Cells via Bioorthogonal Chemical Reporter

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et al. 2022

Angewandte Chemie

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Due to the lack of suitable chemical tools, probing the protein-specific glycation is highly challenging. Herein, we present a strategy based on glycation chemical reporter and proximity-induced FRET signal readout for visualizing protein-specific glycation in living cells. We first developed a bioorthogonal glucose analogue, 6-azido-6-deoxy-D-glucose (6AzGlc), as a novel glycation chemical reporter. Two types of DNA probes, glycation conversion probe and protein targeting probe, were designed to attach to glycation adducts and target proteins, respectively. After the protein was glycated by 6AzGlc, two DNA probes were sequentially applied to the target protein, triggering proximityinduced FRET signal readout. This strategy was successfully used to visualize glucose glycation of several proteins, including PD-L1 and integrin. More importantly, this strategy allowed us to analyze corresponding biological functions of glycated protein in the native environment.

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Visualization of Protein‐Specific Glycation in Living Cells via Bioorthogonal Chemical Reporter

Shao

¹

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Yuan

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Zhou

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et al. 2022

View full text Add to dashboard Cite

Due to the lack of suitable chemical tools, probing the protein-specific glycation is highly challenging. Herein, we present a strategy based on glycation chemical reporter and proximity-induced FRET signal readout for visualizing protein-specific glycation in living cells. We first developed a bioorthogonal glucose analogue, 6-azido-6-deoxy-D-glucose (6AzGlc), as a novel glycation chemical reporter. Two types of DNA probes, glycation conversion probe and protein targeting probe, were designed to attach to glycation adducts and target proteins, respectively. After the protein was glycated by 6AzGlc, two DNA probes were sequentially applied to the target protein, triggering proximityinduced FRET signal readout. This strategy was successfully used to visualize glucose glycation of several proteins, including PD-L1 and integrin. More importantly, this strategy allowed us to analyze corresponding biological functions of glycated protein in the native environment.

show abstract

A Systematic Study of Selective Protein Glycation

Abstract: This is the author manuscript accepted for publication and has undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.

Cited by 7 publications

References 42 publications

Synergistic sequence contributions bias glycation outcomes

Synergistic sequence contributions bias glycation outcomes

Visualization of Protein‐Specific Glycation in Living Cells via Bioorthogonal Chemical Reporter

Visualization of Protein‐Specific Glycation in Living Cells via Bioorthogonal Chemical Reporter

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