Capturing acyl–enzyme intermediates with genetically encoded 2,3-diaminopropionic acid for hydrolase substrate identification

Juan Luo, Yao Yu, Ke Wang, Sizhe He, Longjie Wang, Fangfang Liang, Jason W. Chin, Shan Tang

Published: 2024-06-11 DOI: 10.1038/s41596-024-01006-x

acyl–enzyme intermediates

genetically encoded 2,3-diaminopropionic acid

hydrolase substrates

mass spectrometry

substrate capture

AI 解读

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Heterogeneity（异质性）

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References

Introduction
Materials
Procedure 1: DAP-mediated substrate trapping in mammalian cell lysate
Procedure 2: DAP-mediated substrate trapping in adherent mammalian cells
- Part 1: preparation of DAP-containing UL36USP in HEK293T cells
- Part 2: DAP-mediated substrate trapping in HEK293T cells
Procedure 3: DAP-mediated substrate trapping in suspension mammalian cells
- Part 1: preparation of DAP-containing UL36USP in Expi293 cells
- Part 2: DAP-mediated substrate trapping in Expi293 cells
Procedure 4: substrate identification and validation
- Large-scale preparation of hydrolase–substrate conjugates
- Sample preparation for LC–MS/MS analysis
Troubleshooting
Timing
Anticipated results
- Reporting summary
Acknowledgements

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Capturing acyl–enzyme intermediates with genetically encoded 2,3-diaminopropionic acid for hydrolase substrate identification

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