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Analysis of protein phosphorylation in the regions of consecutive serine/threonine residues by negative ion electrospray collision-induced dissociation. Approach to pinpointing of phosphorylation sites.
Anal Chem. 2007 May 01; 79(9):3476-86.AC

Abstract

Pinpointing of phosphorylation sites by positive ion collision-induced dissociation (CID) in phosphopeptides containing consecutive Ser/Thr residues (Ser/Thr clusters) is frequently hampered by the lack of backbone cleavage between adjacent Ser/Thr or pSer/pThr sites. In this study, we demonstrate that in negative ion collision-induced dissociation phosphorylated and unmodified residues of Ser/Thr clusters exhibit a very selective behavior toward cleavage of their N-Calpha bonds. Ser/Thr clusters were defined as two and more consecutive serine or threonine residues in phosphopeptide sequences. Dissociation reactions at pSer are significantly more abundant than those of unmodified sites. Thr residues exhibit the same effect, but the cleavages occurring at pThr are generally less prominent than those at pSer. The correlation observed between the facility of the amine backbone bond dissociation of phosphopeptides and the presence of the phosphate group on the side chain residues of Ser and Thr is attributed to the different magnitudes of electron density on the Calpha atoms of the amino acid in phosphorylated and unmodified forms. The results of this study indicate that the intensity ratio of the fragments generated by N-Calpha bond cleavage within the phosphopeptide Ser/Thr clusters represents a reliable and general marker for pinpointing of phosphorylation sites. The presented data illustrate that negative ion electrospray CID is superior over the standard positive ion mode approach for the localization of protein phosphorylation inside Ser/Thr clusters.

Authors+Show Affiliations

Central Spectroscopy, German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17388569

Citation

Edelson-Averbukh, Marina, et al. "Analysis of Protein Phosphorylation in the Regions of Consecutive Serine/threonine Residues By Negative Ion Electrospray Collision-induced Dissociation. Approach to Pinpointing of Phosphorylation Sites." Analytical Chemistry, vol. 79, no. 9, 2007, pp. 3476-86.
Edelson-Averbukh M, Pipkorn R, Lehmann WD. Analysis of protein phosphorylation in the regions of consecutive serine/threonine residues by negative ion electrospray collision-induced dissociation. Approach to pinpointing of phosphorylation sites. Anal Chem. 2007;79(9):3476-86.
Edelson-Averbukh, M., Pipkorn, R., & Lehmann, W. D. (2007). Analysis of protein phosphorylation in the regions of consecutive serine/threonine residues by negative ion electrospray collision-induced dissociation. Approach to pinpointing of phosphorylation sites. Analytical Chemistry, 79(9), 3476-86.
Edelson-Averbukh M, Pipkorn R, Lehmann WD. Analysis of Protein Phosphorylation in the Regions of Consecutive Serine/threonine Residues By Negative Ion Electrospray Collision-induced Dissociation. Approach to Pinpointing of Phosphorylation Sites. Anal Chem. 2007 May 1;79(9):3476-86. PubMed PMID: 17388569.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of protein phosphorylation in the regions of consecutive serine/threonine residues by negative ion electrospray collision-induced dissociation. Approach to pinpointing of phosphorylation sites. AU - Edelson-Averbukh,Marina, AU - Pipkorn,Rüdiger, AU - Lehmann,Wolf D, Y1 - 2007/03/28/ PY - 2007/3/29/pubmed PY - 2007/6/15/medline PY - 2007/3/29/entrez SP - 3476 EP - 86 JF - Analytical chemistry JO - Anal Chem VL - 79 IS - 9 N2 - Pinpointing of phosphorylation sites by positive ion collision-induced dissociation (CID) in phosphopeptides containing consecutive Ser/Thr residues (Ser/Thr clusters) is frequently hampered by the lack of backbone cleavage between adjacent Ser/Thr or pSer/pThr sites. In this study, we demonstrate that in negative ion collision-induced dissociation phosphorylated and unmodified residues of Ser/Thr clusters exhibit a very selective behavior toward cleavage of their N-Calpha bonds. Ser/Thr clusters were defined as two and more consecutive serine or threonine residues in phosphopeptide sequences. Dissociation reactions at pSer are significantly more abundant than those of unmodified sites. Thr residues exhibit the same effect, but the cleavages occurring at pThr are generally less prominent than those at pSer. The correlation observed between the facility of the amine backbone bond dissociation of phosphopeptides and the presence of the phosphate group on the side chain residues of Ser and Thr is attributed to the different magnitudes of electron density on the Calpha atoms of the amino acid in phosphorylated and unmodified forms. The results of this study indicate that the intensity ratio of the fragments generated by N-Calpha bond cleavage within the phosphopeptide Ser/Thr clusters represents a reliable and general marker for pinpointing of phosphorylation sites. The presented data illustrate that negative ion electrospray CID is superior over the standard positive ion mode approach for the localization of protein phosphorylation inside Ser/Thr clusters. SN - 0003-2700 UR - https://www.unboundmedicine.com/medline/citation/17388569/Analysis_of_protein_phosphorylation_in_the_regions_of_consecutive_serine/threonine_residues_by_negative_ion_electrospray_collision_induced_dissociation__Approach_to_pinpointing_of_phosphorylation_sites_ DB - PRIME DP - Unbound Medicine ER -