Histone Synthesis

Chatterjee, Champak

doi:10.1002/9783527823567.ch18

Cited by 1 publication

(1 citation statement)

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“…In addition, many ncAAs of interest are not commercially available and therefore often a multi-step synthetic process is required to yield the desired ncAAs beforehand. Another strategy, exemplified primarily by thioester-mediated native chemical ligation or expressed protein ligation, can produce full-length histone proteins with a wider range of relevant PTMs 9,10 when paired with peptide synthesis to introduce a site-specifically modified segment via 'linear', backbone-amide ligation. However, production of full-length proteins in this way may be time intensive and necessitates peptide chemistry knowledge.…”

Section: Introductionmentioning

confidence: 99%

Posttranslational, site-directed photochemical fluorine editing of protein sidechains to probe residue oxidation state via 19F-nuclear magnetic resonance

et al. 2023

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The fluorination of amino acid residues represents a near-isosteric alteration with the potential to report on biological pathways, yet the site-directed editing of carbon-hydrogen (C-H) bonds in complex biomolecules to carbon-fluorine (C-F) ones is challenging, resulting in its limited exploitation. Here, we describe a protocol for the post-translational and site-directed alteration of native γCH2 to γCF2 in protein sidechains. This alteration allows the installation of difluorinated sidechain analogues of proteinogenic amino acids, in both native and modified states. This chemical editing is robust, mild, fast, and highly efficient, exploiting photochemical and radicalmediated C-C-bonds grafted onto easy-to-access cysteine-derived dehydroalanine-containing proteins as starting materials. The heteroaryl-sulfonyl 'pySOOF' reagent required for generating the key carbon-centred C• radicals that install the sidechain can be synthesized in two-to-six steps from commercially available precursors. This workflow allows the non-expert to create fluorinated proteins within 24 hours, starting from a corresponding purified cysteine-containing protein precursor, without the need for bespoke biological systems. As an example, we readily introduce three γCF2-containing methionines in all three progressive oxidation states (sulfide, sulfoxide and sulfone) as D-/L-forms into Histone eH3.1 at site 4 (a relevant lysine to methionine oncomutation site) and each can be detected by 19 F-nuclear magnetic resonance of the γCF2 group, as well as the two diastereomers of the sulfoxide, even when found in a complex protein mixture of all three.The site-directed editing of C-HC-F enables the use of γCF2 as a highly-sensitive, 'zero-sizezero-background' label in protein sidechains, which may be used to probe biological phenomena, protein structures and/or protein-ligand interactions by 19 F-based detection methods.

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Section: Introductionmentioning

confidence: 99%