Bioinspired Thiophosphorodichloridate Reagents for Chemoselective Histidine Bioconjugation

Abstract

Site-selective bioconjugation to native protein residues is a powerful tool for protein functionalization, with cysteine and lysine side chains being the most common points for attachment owing to their high nucleophilicity. We now report a strategy for histidine modification using thiophosphorodichloridate reagents that mimic post-translational histidine phosphorylation, enabling fast and selective labeling of protein histidines under mild conditions where various payloads can be introduced via copper-assisted alkyne-azide cycloaddition (CuAAC) chemistry. We establish that these reagents are particularly effective at covalent modification of His-tags, which are common motifs to facilitate protein purification, as illustrated by selective attachment of polyarginine cargoes to enhance the uptake of proteins into living cells. This work provides a starting point for probing and enhancing protein function using histidine-directed chemistry.

CRISPR ‐ TAPE : protein‐centric CRISPR guide design for targeted proteome engineering

“…The profiling of amino acid reactivity within proteins is a rapidly expanding field, with this chemical proteomic strategy having been successfully applied to profile the reactivity of serine (Kidd et al , ), cysteine (Weerapana et al , ), lysine (Hacker et al , ), histidine (Jia et al , ), tyrosine (Hahm et al , ) and methionine residues (Ohata et al , ). A method to efficiently prioritize these reactive sites according to their contribution to protein function would optimize pipelines for target‐based screening platforms.…”

Aktivitätsbasierte Sensorik: ein synthetisch‐methodischer Ansatz für die selektive molekulare Bildgebung und darüber hinaus.

“…[323][324][325][326][327][328][329] Richtungweisend dafüri st die Entwicklung einer Chemoproteomikplattform zur Identifizierung hyperreaktiver Cysteine durch isoTOP-ABPP (isotopic Tandem Orthogonal Proteolysis -A ctivity-Based Protein Profiling). [330] Aufbauend auf umfangreichen ABPP-Arbeiten über Cystein-, [331,332] Lysin- [333,334] und Serinnachweise [335] haben wir und andere Biokonjugationsstrategien fürw eitere native Aminosäuren verfolgt, darunter Methionin, [336][337][338] Histidin, [339] Ty rosin [340][341][342] und Tryptophan. [343] So haben unser Labor und Tostes Arbeitsgruppe gemeinsam eine neue ABS-Methode fürd ie Methionin-Biokonjugation entwickelt, genannt ReACT (Redox-Activated Chemical Tagging), bei der wir die nativen Redoxeigenschaften der Oxidation von Methionin zu Methioninsulfoxid nutzen, um Oxaziridinreagenzien zu entwickeln, die die isoelektronische Nitrenübertragung zu einem Sulfimidaddukt auslçsen.…”

Proximity‐driven, Regioselective Chemical Modification of Peptides and Proteins

“…These efforts have been extensively reviewed. [1][2][3][4][5] More recently, several more of these methods have been reported that utilize novel functional groups to demonstrate impressive chemoselectivity to Cys, [6][7][8][9] Lys, [10][11][12][13] His, [14][15][16] Met, [17][18][19] Trp, [20] and Tyr. [21] Relatedly, dehydroalanine (Dha)-mediated modification is an indirect but notable approach.…”