A New Derivatization Strategy for the Analysis of Phosphopeptides by Precursor Ion Scanning in Positive Ion Mode

Overview

Journal J Am Soc Mass Spectrom

Specialty Chemistry

Date 2002 Sep 10

PMID 12216740

Citations 14

Authors

Hanno Steen

Matthias Mann

Affiliations

Soon will be listed here.

Abstract

Although numerous strategies have been devised to analyze protein phosphorylation, an abundant intracellular protein modification, there is still a need for different methods for the analysis of this modification. A method to both detect and localize the phosphorylation within a protein/peptide is especially required. In this paper, a new strategy is described, which makes use of beta-elimination/Michael addition reactions to introduce a functional group at the original site of phosphorylation, which gives rise to a dimethylamine-containing sulfenic acid derivative with a unique m/z value. This enables the detection of the phosphorylated species within peptide mixtures by sensitive and specific precursor ion scanning in positive ion mode. Working under acidic conditions in positive ion mode has the added advantage that subsequent normal peptide sequencing for the exact localization can be performed. No other peptide derived fragment ion is observed at the m/z value of the sulfenic acid derivative formed, thus specific precursor ion experiments can also be carried out on instruments with low fragment ion resolution and lends itself to LC-MS/MS approaches when skimmer fragmentation routines or triple quadrupole mass spectrometers are used.

Citing Articles

Selective Enrichment of Histidine Phosphorylated Peptides Using Molecularly Imprinted Polymers.

Incel A, Diez I, Wierzbicka C, Gajoch K, Jensen O, Sellergren B Anal Chem. 2021; 93(8):3857-3866.

PMID: 33591162 PMC: 8023515. DOI: 10.1021/acs.analchem.0c04474.

Hydroxyphenylation of Histone Lysines: Post-translational Modification by Quinone Imines.

Ravindra K, Trudel L, Wishnok J, Wogan G, Tannenbaum S, Skipper P ACS Chem Biol. 2016; 11(5):1230-7.

PMID: 26866676 PMC: 5495105. DOI: 10.1021/acschembio.5b00923.

Study of phosphorylation events for cancer diagnoses and treatment.

Lopez Villar E, Madero L, A Lopez-Pascual J, Cho W Clin Transl Med. 2015; 4(1):59.

PMID: 26055493 PMC: 4460185. DOI: 10.1186/s40169-015-0059-0.

Functional phosphoproteomic mass spectrometry-based approaches.

Lopez E, Wang X, Madero L, Lopez-Pascual J, Latterich M Clin Transl Med. 2013; 1(1):20.

PMID: 23369623 PMC: 3560980. DOI: 10.1186/2001-1326-1-20.

Relevant phosphoproteomic and mass spectrometry: approaches useful in clinical research.

Lopez E, Rodriguez Munoz S, Pascual J, Madero L Clin Transl Med. 2013; 1(1):2.

PMID: 23369602 PMC: 3552569. DOI: 10.1186/2001-1326-1-2.

References

Resing K, Johnson R, Walsh K . Mass spectrometric analysis of 21 phosphorylation sites in the internal repeat of rat profilaggrin, precursor of an intermediate filament associated protein. Biochemistry. 1995; 34(29):9477-87. DOI: 10.1021/bi00029a024. View

Steen H, Kuster B, Fernandez M, Pandey A, Mann M . Detection of tyrosine phosphorylated peptides by precursor ion scanning quadrupole TOF mass spectrometry in positive ion mode. Anal Chem. 2001; 73(7):1440-8. DOI: 10.1021/ac001318c. View

Wilm M, Neubauer G, Mann M . Parent ion scans of unseparated peptide mixtures. Anal Chem. 1996; 68(3):527-33. DOI: 10.1021/ac950875+. View

Quadroni M, James P, Carafoli E . Isolation of phosphorylated calmodulin from rat liver and identification of the in vivo phosphorylation sites. J Biol Chem. 1994; 269(23):16116-22. View

Meyer H, Hoffmann-Posorske E, Korte H, Heilmeyer Jr L . Sequence analysis of phosphoserine-containing peptides. Modification for picomolar sensitivity. FEBS Lett. 1986; 204(1):61-6. DOI: 10.1016/0014-5793(86)81388-6. View

Cramer R, Corless S . The nature of collision-induced dissociation processes of doubly protonated peptides: comparative study for the future use of matrix-assisted laser desorption/ionization on a hybrid quadrupole time-of-flight mass spectrometer in proteomics. Rapid Commun Mass Spectrom. 2001; 15(22):2058-66. DOI: 10.1002/rcm.485. View

Annan R, Huddleston M, Verma R, Deshaies R, Carr S . A multidimensional electrospray MS-based approach to phosphopeptide mapping. Anal Chem. 2001; 73(3):393-404. DOI: 10.1021/ac001130t. View

Shevchenko A, Wilm M, Vorm O, Mann M . Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 1996; 68(5):850-8. DOI: 10.1021/ac950914h. View

Neubauer G, Mann M . Mapping of phosphorylation sites of gel-isolated proteins by nanoelectrospray tandem mass spectrometry: potentials and limitations. Anal Chem. 1999; 71(1):235-42. DOI: 10.1021/ac9804902. View

10.

Jaffe H, Veeranna , Pant H . Characterization of serine and threonine phosphorylation sites in beta-elimination/ethanethiol addition-modified proteins by electrospray tandem mass spectrometry and database searching. Biochemistry. 1998; 37(46):16211-24. DOI: 10.1021/bi981264p. View

11.

Weckwerth W, Willmitzer L, Fiehn O . Comparative quantification and identification of phosphoproteins using stable isotope labeling and liquid chromatography/mass spectrometry. Rapid Commun Mass Spectrom. 2000; 14(18):1677-81. DOI: 10.1002/1097-0231(20000930)14:18<1677::AID-RCM84>3.0.CO;2-N. View

12.

Goshe M, Conrads T, Panisko E, Angell N, Veenstra T, Smith R . Phosphoprotein isotope-coded affinity tag approach for isolating and quantitating phosphopeptides in proteome-wide analyses. Anal Chem. 2001; 73(11):2578-86. DOI: 10.1021/ac010081x. View

13.

Nuwaysir L, Stults J . Electrospray ionization mass spectrometry of phosphopeptides isolated by on-line immobilized metal-ion affinity chromatography. J Am Soc Mass Spectrom. 2013; 4(8):662-9. DOI: 10.1016/1044-0305(93)85031-R. View

14.

Steen H, Kuster B, Mann M . Quadrupole time-of-flight versus triple-quadrupole mass spectrometry for the determination of phosphopeptides by precursor ion scanning. J Mass Spectrom. 2001; 36(7):782-90. DOI: 10.1002/jms.174. View

15.

Steen H, Mann M . Similarity between condensed phase and gas phase chemistry: fragmentation of peptides containing oxidized cysteine residues and its implications for proteomics. J Am Soc Mass Spectrom. 2001; 12(2):228-32. DOI: 10.1016/S1044-0305(00)00219-1. View

16.

Polce M, Ren D, Wesdemiotis C . Dissociation of the peptide bond in protonated peptides. J Mass Spectrom. 2001; 35(12):1391-8. DOI: 10.1002/1096-9888(200012)35:12<1391::AID-JMS85>3.0.CO;2-1. View

17.

Gobom J, Nordhoff E, Mirgorodskaya E, Ekman R, Roepstorff P . Sample purification and preparation technique based on nano-scale reversed-phase columns for the sensitive analysis of complex peptide mixtures by matrix-assisted laser desorption/ionization mass spectrometry. J Mass Spectrom. 1999; 34(2):105-16. DOI: 10.1002/(SICI)1096-9888(199902)34:2<105::AID-JMS768>3.0.CO;2-4. View

18.

Carr S, Huddleston M, Annan R . Selective detection and sequencing of phosphopeptides at the femtomole level by mass spectrometry. Anal Biochem. 1996; 239(2):180-92. DOI: 10.1006/abio.1996.0313. View

19.

Wysocki V, Tsaprailis G, Smith L, Breci L . Mobile and localized protons: a framework for understanding peptide dissociation. J Mass Spectrom. 2001; 35(12):1399-406. DOI: 10.1002/1096-9888(200012)35:12<1399::AID-JMS86>3.0.CO;2-R. View

20.

Stensballe A, Andersen S, Jensen O . Characterization of phosphoproteins from electrophoretic gels by nanoscale Fe(III) affinity chromatography with off-line mass spectrometry analysis. Proteomics. 2001; 1(2):207-22. DOI: 10.1002/1615-9861(200102)1:2<207::AID-PROT207>3.0.CO;2-3. View