Labeling Preferences of Diazirines with Protein Biomolecules
| Autor: | Andrew C. Huang, Lyn H. Jones, Giovanni Muncipinto, Christina M. Woo, Hung-Yi Wu, Matthew T Labenski, Alexander V West |
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| Rok vydání: | 2021 |
| Předmět: |
Proteome
Protein Conformation Photoaffinity Labels 010402 general chemistry 01 natural sciences Biochemistry Catalysis chemistry.chemical_compound Colloid and Surface Chemistry Protein structure Humans Reactivity (chemistry) Amino Acids Alkyl chemistry.chemical_classification Binding Sites Photoaffinity labeling Chemistry Biomolecule Voltage-Dependent Anion Channel 1 Proteins General Chemistry Diazonium Compounds Hydrogen-Ion Concentration Combinatorial chemistry 0104 chemical sciences Amino acid Diazomethane Diazirine Diazo |
| Zdroj: | Journal of the American Chemical Society. 143(17) |
| ISSN: | 1520-5126 |
| Popis: | Diazirines are widely used in photoaffinity labeling (PAL) to trap noncovalent interactions with biomolecules. However, design and interpretation of PAL experiments is challenging without a molecular understanding of the reactivity of diazirines with protein biomolecules. Herein, we report a systematic evaluation of the labeling preferences of alkyl and aryl diazirines with individual amino acids, single proteins, and in the whole cell proteome. We find that alkyl diazirines exhibit preferential labeling of acidic amino acids in a pH-dependent manner that is characteristic of a reactive alkyl diazo intermediate, while the aryl-fluorodiazirine labeling pattern reflects reaction primarily through a carbene intermediate. From a survey of 32 alkyl diazirine probes, we use this reactivity profile to rationalize why alkyl diazirine probes preferentially enrich highly acidic proteins or those embedded in membranes and why probes with a net positive charge tend to produce higher labeling yields in cells and in vitro. These results indicate that alkyl diazirines are an especially effective chemistry for surveying the membrane proteome and will facilitate design and interpretation of biomolecular labeling experiments with diazirines. |
| Databáze: | OpenAIRE |
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