4-Phosphopyrazol-2-yl alanine: a non-hydrolysable analogue of phosphohistidine

Lilley, Matthew; Mambwe, Bezaleel; Thompson, Mark J.; Jackson, Richard F. W.; Muimo, Richmond

doi:10.1039/c5cc01811k

Cited by 30 publications

(27 citation statements)

References 28 publications

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“…However, a growing body of evidence (Besant & Attwood, ; Fan et al , ; Fraczyk et al , ; Fuhs et al , ; Shen et al , ; Wieland & Attwood, ; Panda et al , ; Srivastava et al , ; Xu et al , ,b; Fuhs & Hunter, ) indicates that phosphorylation of other amino acids, termed here “non‐canonical” phosphorylation, including His, Asp, Glu, Lys, Arg and Cys [in addition to pyrophosphorylation of Ser and Thr to form ppSer and ppThr (Chanduri et al , ; Harmel & Fiedler, ), and Lys polyphosphorylation (Bentley‐DeSousa & Downey, )], may also regulate protein signalling functions. Of note, the generation of both site‐specific and generic antibodies against phosphohistidine (pHis) (Kee et al , , ; Fuhs et al , ; Lilley et al , ) has recently allowed mammalian cell‐type independent roles for this PTM to be elucidated (Fuhs et al , ) in processes as diverse as ion channel regulation (Srivastava et al , ) and T‐cell signalling (Panda et al , ), and during cell proliferation, differentiation and migration. In contrast, and despite occasional reports in the literature (Besant et al , ; Attwood et al , ), the prevalence (or otherwise) of other non‐canonical protein phosphorylation events has not been reliably confirmed in human cells, and so specific information about the extent and details of the sites of modification is lacking.…”

Section: Introductionmentioning

confidence: 99%

Strong anion exchange‐mediated phosphoproteomics reveals extensive human non‐canonical phosphorylation

et al. 2019

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Phosphorylation is a key regulator of protein function under (patho)physiological conditions, and defining site‐specific phosphorylation is essential to understand basic and disease biology. In vertebrates, the investigative focus has primarily been on serine, threonine and tyrosine phosphorylation, but mounting evidence suggests that phosphorylation of other “non‐canonical” amino acids also regulates critical aspects of cell biology. However, standard methods of phosphoprotein characterisation are largely unsuitable for the analysis of non‐canonical phosphorylation due to their relative instability under acidic conditions and/or elevated temperature. Consequently, the complete landscape of phosphorylation remains unexplored. Here, we report an unbiased phosphopeptide enrichment strategy based on strong anion exchange (SAX) chromatography (UPAX), which permits identification of histidine (His), arginine (Arg), lysine (Lys), aspartate (Asp), glutamate (Glu) and cysteine (Cys) phosphorylation sites on human proteins by mass spectrometry‐based phosphoproteomics. Remarkably, under basal conditions, and having accounted for false site localisation probabilities, the number of unique non‐canonical phosphosites is approximately one‐third of the number of observed canonical phosphosites. Our resource reveals the previously unappreciated diversity of protein phosphorylation in human cells, and opens up avenues for high‐throughput exploration of non‐canonical phosphorylation in all organisms.

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Section: Introductionmentioning

confidence: 99%

Strong anion exchange‐mediated phosphoproteomics reveals extensive human non‐canonical phosphorylation

et al. 2019

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“…These polyclonal antibodies were also found to have strong selectivity for pHis over pTyr, on dot blots, immunoblots and ELISA against BSA phosphorylated aminoacid conjugates as standards. Polyclonal antibodies reported by Kee et al 103 were used to detect various in vitro His phosphorylated proteins including PGAM1, mammalian histone H4 and PtsI, whereas those by Lilley et al 111 antibodies were used to detect immunoprecipitated protein Gβ and both NDPK-A/B from HBE cells.…”

Section: Phosphohistidine Analogues and Antibodiesmentioning

confidence: 99%

Advances in development of new tools for the study of phosphohistidine

Makwana

Muimo

Jackson

2018

Laboratory Investigation

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Protein phosphorylation is an important post-translational modification that is an integral part of cellular function. The O-phosphorylated amino-acid residues, such as phosphoserine (pSer), phosphothreonine (pThr) and phosphotyrosine (pTyr), have dominated the literature while the acid labile N-linked phosphorylated amino acids, such as phosphohistidine (pHis), have largely been historically overlooked because of the acidic conditions routinely used in amino-acid detection and analysis. This review highlights some misinterpretations that have arisen in the existing literature, pinpoints outstanding questions and potential future directions to clarify the role of pHis in mammalian signalling systems. Particular emphasis is placed on pHis isomerization and the hybrid functionality for both pHis and pTyr of the proposed τ-pHis analogue bearing the triazole residue.

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“…Synthetic nonhydrolyzable analogues of pHis [105][106][107][108][109] and pArg [110,111] have been extremely useful as haptens to generate pHis-or pArg-specific antibodies, whichh ave been used in the detection and enrichmento ft hese PTM-bearing proteins (Scheme 5). [13,28,30] These analogues replaced the labile phosphoramidate with much more stable phosphonate or sulfonate.…”

Section: Chemical Toolsmentioning

confidence: 99%

Recent Updates on Protein N‐Phosphoramidate Hydrolases

2018

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In contrast to well‐recognized protein phosphorylation on the side‐chain oxygen of Ser, Thr, or Tyr residues, analogous phosphoramidation of the nitrogen of His, Lys, and Arg side chains remains much less investigated, mainly due to the instability of post‐translational modifications and technical difficulties involved in their analysis. For example, reports on the enzyme activities responsible for the formation and hydrolysis of these phosphoramidates date back to as early as the 1950s, but some of these enzymes have only recently been identified and functionally characterized; this has been aided by the development of novel research tools. In this review, we summarize current knowledge of the enzymes that hydrolyze protein N‐phosphoramidates, in terms of their structure, activities, and biological functions, as well as the chemical tools used to investigate them.

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4-Phosphopyrazol-2-yl alanine: a non-hydrolysable analogue of phosphohistidine

Cited by 30 publications

References 28 publications

Strong anion exchange‐mediated phosphoproteomics reveals extensive human non‐canonical phosphorylation

Strong anion exchange‐mediated phosphoproteomics reveals extensive human non‐canonical phosphorylation

Advances in development of new tools for the study of phosphohistidine

Recent Updates on Protein N‐Phosphoramidate Hydrolases

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