Synthesis and characterization of histidine‐phosphorylated peptides

Medzihradszky, Katalin F.; Phillipps, Nancy J.; Senderowicz, Lionel; Wang, Ping; Turck, Christoph W.

doi:10.1002/pro.5560060704

Cited by 60 publications

(58 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus far the only observation of extensive gas-phase dephosphorylation during ESI involved the highly reactive phospho-histidine species (35). Therefore, the observations described above appear to be diagnostic for the presence of a sulfono-moiety (36).…”

Section: Resultsmentioning

confidence: 89%

O-Sulfonation of Serine and Threonine

Medzihradszky

Darula

Perlson

et al. 2004

Molecular & Cellular Proteomics

130

View full text Add to dashboard Cite

Protein sulfonation on serine and threonine residues is described for the first time. This post-translational modification is shown to occur in proteins isolated from organisms representing a broad span of eukaryote evolution, including the invertebrate mollusk Lymnaea stagnalis, the unicellular malaria parasite Plasmodium falciparum, and humans. Detection and structural characterization of this novel post-translational modification was carried out using liquid chromatography coupled to electrospray tandem mass spectrometry on proteins including a neuronal intermediate filament and a myosin light chain from the snail, a cathepsin-C-like enzyme from the parasite, and the cytoplasmic domain of the human orphan receptor tyrosine kinase Ror-2. These findings suggest that sulfonation of serine and threonine may be involved in multiple functions including protein assembly and signal transduction. Molecular & Cellular Proteomics 3:429 -443, 2004.Sulfonation occurs as a common enzymatic modification of endogenous substances including proteins, carbohydrates, catecholamines, and estrogenic steroids as well as xenobiotic chemicals (1). Sulfonation refers to the transfer of the sulfonate group (SO 3 Ϫ1 ) from 3Ј-phosphoadenosine-5Ј-phosphosulfate (PAPS), 1 the only known sulfonate donor (2), and can occur through several types of linkages, such as esters and anhydrides (O-sulfonation), amides (N-sulfonation), and thioesters (S-sulfonation), of which O-sulfonation is the most prominent (3). The transfer of SO 3 Ϫ1 to a hydroxyl or phenolic acceptor (O-sulfonation) generates a sulfono-derivative, and this reaction has commonly been referred to as sulfation rather than the more accurate O-sulfonation. The majority of cellular sulfonation is of the O type and occurs primarily on polysaccharides, steroids, catecholamines, and thyroid hormones (1). These reactions are catalyzed by the soluble cytosolic sulfotransferases and appear to alter their bioactivity. For example, estrogen, testosterone, and thyroid hormones (T 3 and T 4 ) can interact with their respective receptors to regulate transcription, whereas their sulfate-containing moieties cannot. Furthermore, the half-life of these compounds in blood is significantly shorter than that of their conjugated counterparts, suggesting that sulfonation maintains these compounds in an inactive state ready for rapid deployment by the removal of the sulfonyl group.While the cytosolic sulfotransferases conjugate cell-permeable or intracellular compounds, the membrane-bound Golgiassociated sulfotransferases are primarily responsible for sulfonation of extracellular proteins via a co-or post-translational mechanism. The membrane-bound sulfotransferases are responsible for the sulfonation of various glycosaminoglycans, such as heparin and heparan sulfate. Additionally, these enzymes catalyze the direct sulfonation of proteins on the O 4 position of tyrosine residues (4). It is one of the last modifications to occur during protein transiting the trans-Golgi and thus has been found almost ex...

show abstract

Section: Resultsmentioning

confidence: 89%

O-Sulfonation of Serine and Threonine

Medzihradszky

Darula

Perlson

et al. 2004

Molecular & Cellular Proteomics

130

View full text Add to dashboard Cite

show abstract

“…1!. The histidine phosphorylation of the peptide was carried out with potassium phosphoramidate as previously described~Wei & Matthews, 1991;Medzihradszky et al, 1997;Senderowicz et al, 1997!. For the preparation of the histidine-thiophosphorylated peptide, we developed a procedure that employs PSCl 3 as a phosphorylation reagent. We had to resort to this rather reactive compound because all attempts to prepare potassium thiophosphoramidate for use as the thiophosphorylating reagent failed.…”

Section: Resultsmentioning

confidence: 99%

“…For histidine phosphorylation, 5 mg of peptide was dissolved in 200 mL 10 mM ammonium bicarbonate and the pH adjusted to 8 with 6 N KOH. To this peptide solution, 50 mg of potassium phosphoramidate~Wei & Matthews, 1995;Medzihradszky et al, 1997! was added and the mixture reacted overnight at RT~20 8C!.…”

Section: Peptidesmentioning

confidence: 99%

Protein histidine phosphorylation: Increased stability of thiophosphohistidine

et al. 1999

Self Cite

View full text Add to dashboard Cite

Posttranslational phosphorylation of proteins is an important event in many cellular processes. Whereas phosphoesters of serine, threonine and tyrosine have been extensively studied, only limited information is available for other amino acids modified by a phosphate group. The formation of phosphohistidine residues in proteins has been discovered in prokaryotic organisms as well as in eukaryotic cells. The ability to biochemically analyze phosphohistidine residues in proteins, however, is severely hampered by its extreme lability under acidic conditions. In our studies we have found that by replacing the phosphate linked to the histidine residue with a thiophosphate, a phosphohistidine derivative with increased stability is formed. This allows the analysis of phosphohistidine-containing proteins by established biochemical techniques and will greatly aid in the investigation of the role of this posttranslational modification in cellular processes.

show abstract

“…However, reaction conditions have to be chosen carefully to avoid both incomplete esterification and side reactions for instance with asparagines because they increase sample complexity and interfere with subsequent mass spectrometric analysis. Thus, various metal-ions such as Fe 31 , Ga 31 , Al 31 or Zr 41 have been used for better selectivity and phosphopeptide recovery [26,27]. Especially Ga 31 -ions have proven well [28] in different studies but Fe 31 -based methods are still used more often.…”

Section: Detection Of Phosphoproteins In 2-d Gelsmentioning

confidence: 99%

“…Phosphorylations that are instable under all described conditions are mostly analyzable exclusively by indirect methods or techniques with rather poor detection limits such as HPLC followed by dephosphorylation of the peptide and subsequent amino acid analysis or NMR-measurements, etc. [1,41].…”

Section: Identification Of Phosphorylated Amino Acid Residuesmentioning

confidence: 99%

State-of-the-art in phosphoproteomics

2005

View full text Add to dashboard Cite

Presently, phosphorylation of proteins is the most studied and best understood PTM. However, the analysis of phosphoproteins and phosphopeptides is still one of the most challenging tasks in contemporary proteome research. Since not every phosphoprotein is accessible by a certain method and identification of the phosphorylated amino acid residue is required in the majority of cases, various strategies for the detection and localization of phosphorylations have been developed. Identification and localization of protein phosphorylations is mostly done by MS nowadays but phosphoproteins and -peptides are often suppressed in comparison to the unphosphorylated species if measured in complex mixtures. Thus, the isolation of pure phosphopeptide samples is a main task. This review gives an overview over the most frequently used methods in isolation and detection of phosphoproteins and -peptides such as specific enrichment or separation strategies as well as the localization of the phosphorylated residues by various mass spectrometric techniques.

show abstract

Synthesis and characterization of histidine‐phosphorylated peptides

Cited by 60 publications

References 22 publications

O-Sulfonation of Serine and Threonine

O-Sulfonation of Serine and Threonine

Protein histidine phosphorylation: Increased stability of thiophosphohistidine

State-of-the-art in phosphoproteomics

Contact Info

Product

Resources

About