Coupling capillary zone electrophoresis with electron. Similar to electron capture dissociation, etd induces fragmentation of large, multiplycharged cations by transferring electrons to them. The mechanisms of electron capture and electron transfer dissociation ecd and etd are investigated by covalently attaching a freeradical hydrogen atom. Electron transfer dissociation mass spectrometry in. Since its discovery in 1998, electron capture dissociation ecd has been broadly applied to the structural characterization of a wide variety of biomolecules, including proteinspeptides, dnasrnas, carbohydrates, lipids, and protein complexes. Pdf structural analysis of intact monoclonal antibodies. Us20110062323a1 electron transfer dissociation device. The pairing dissociation energy and electron attracting ability of each anionic structure are predicted and compared with those of the neutral gc and the hydrogenated gc base pairs. Peptide and protein sequence analysis by electron transfer. Tunable infrared freeelectron lasers fels, such as felix at our institute, have played an important role in the recent development of iris, and.
Nov 15, 2015 electron capture dissociation ecd and electron transfer dissociation etd are invaluable tools for elucidating structures of peptides and proteins in mass spectrometry msbased proteomics. Etd capability can be added to many thermo scientific ion. A control system determines the degree of fragmentation and charge reduction of precursor ions within the ion guide and varies the speed at which ions are transmitted through the ion guide in order to optimise the fragmentation and charge reduction process. Electron transfer dissociation etd of peptides containing. Electron transfer dissociation of peptide anions sciencedirect. In etd, one uses collisions with a molecular anion a to transfer an electron to the peptide or protein cation. Electrontransfer dissociation etd is a variation of ecd, in which an ionion reaction, for example, between an anthracene anion and a multiprotonated peptide cation, is used to transfer an electron to the peptide ion. Electron transfer dissociation of modified peptides and. Apr 14, 2006 we demonstrated how the subcycle evolution of the electric field of light can be used to control the motion of bound electrons. Peptide sequence analysis using a combination of gasphase ionion chemistry and tandem mass spectrometry msms is demonstrated.
Capture of a free electron by an ion in the gas phase can be. The nonadiabatic electron transfer between donor d and acceptor a centers is treated by the fgr equation 1. Results are presented for the dissociative ionization of deuterium molecules d2. Here are links to additional resources that might be useful in interpreting etd spectra.
Electron capture dissociation ecd and electron transfer dissociation etd are invaluable tools for elucidating structures of peptides and proteins in mass spectrometry msbased proteomics. Aug 28, 2015 electron transfer dissociation is a popular peptide fragmentation technique in mass spectrometry. In the last decade, researchers have developed an alternative, electron capture dissociation ecd, which involves reacting multiplyp. In contrast, the etd spectrum displays a rich fragmentation pattern of c and z ions that reveals the exact location of the phosphorylation t. Halogenbond assisted photoinduced electron transfer. Sulfopeptide fragmentation in electroncapture and electron. The development of electron based, unimolecular dissociation mass spectrometric methods, i. Jun 29, 2004 peptide sequence analysis using a combination of gasphase ionion chemistry and tandem mass spectrometry msms is demonstrated.
Traditional electrontransfer dissociation etd experiments operate through a. In all, we identified a total of 1,435 phosphorylation sites from human embryonic kidney 293t cells, of which. The cid spectrum shows a few b and y ions, but is dominated by a peak corresponding to the loss of phosphoric acid. Leveraging electron transfer dissociation for site selective radical. Apr 20, 2016 electron capture dissociation ecd and electron transfer dissociation etd in metalpeptide complexes are dependent on the metal cation in the complex. Ionion reactions of multiply deprotonated peptide anions with xenon radical cations result in electron abstraction to generate chargereduced peptide anions containing a freeradical site. We demonstrated how the subcycle evolution of the electric field of light can be used to control the motion of bound electrons. We use the method to describe the electrontransfer induced dissociation of the mchlorocyanobenzene radical anion in aqueous solution. Electrontransfer dissociation an overview sciencedirect.
It is one of the most widely used techniques for activation and dissociation of mass selected precursor ion in msms. Electron transfer dissociation of peptide anions request pdf. Singly charged anthracene anions transfer an electron to multiply protonated peptides in a radio frequency quadrupole linear ion trap qlt and induce fragmentation of the peptide backbone along pathways that are analogous to those observed in electron capture. Electron transfer dissociation is a supremely useful tool for the analysis of posttranslationally modified proteins. Ionion reactions of multiply deprotonated peptide anions with xenon radical cations result in electron abstraction to generate chargereduced peptide anions containing a free radical site. Electron transfer dissociation etd is a variation of ecd, in which an ionion reaction, for example, between an anthracene anion and a multiprotonated peptide cation, is used to transfer an electron to the peptide ion. Investigation of the mechanism of electron capture and. The tutorial is designed to teach you how to interpret and identify etd spectra manually. Etd allows for rapid peptide sequencing, with speeds similar to ion trap cad, but is operated such that ions of mass lower than 100 mz are detected regardless of precursor mz i.
Electron transfer dissociation mass spectrometry in proteomics kim, min. Radical anions for use in the fragmentation of positively charged biopolymer ions by means of electron transfer are produced from substances previously unknown for use as etd production substances. Structural analysis of intact monoclonal antibodies by electron transfer dissociation mass spectrometry. Where bdfe is the bond dissociation free energy for the rx bond. Structural identification of electron transfer dissociation products in. Electron transfer dissociation is complementary to cad and can be especially useful for sequencing peptides containing posttranslational modification ptm 28 36.
The effect of charge state on the dissociation of polypeptides upon electron transfer from a radical anion was investigated by reacting azobenzene anions with different charge states of peptides including tryptic peptides of myoglobin, model peptides, and peptides of relatively large size. Collisionactivated dissociation cad, the most widely used peptide ion fragmentation technique for peptide sequence analysis by tandem mass spectrometry, works great for small peptides but is problematic for labile posttranslational modifications ptms. May 01, 2008 collisionactivated dissociation cad, the most widely used peptide ion fragmentation technique for peptide sequence analysis by tandem mass spectrometry, works great for small peptides but is problematic for labile posttranslational modifications ptms. Protoncoupled electron transfer with metal complexes. Fundamentals of electron capture dissociation fourier. The ultrafast decay of this excited complex to the ground state was ascribed to an electron transfer quenching by the xb donors. The technique that satisfies these two requirements for anion tandem mass spectrometry is negative electron transfer dissociation netd 29, 30, the negativeion complement of electron transfer. Electron transfer dissociation is a popular peptide fragmentation technique in mass spectrometry. A perspective on maldi alternatives total solventfree. Charge remote fragmentation in electron capture and electron.
Etd reactions resulting from electron transfer, a type of ion chemistry, include. In all, we identified a total of 1,435 phosphorylation sites from human embryonic kidney 293t cells, of which 1,141. Electron capture dissociation ecd is a method of fragmenting gasphase ions for structure elucidation of peptides and proteins in tandem mass spectrometry. Electron transfer dissociation last updated july 27, 2019 an ion trap mass spectrometer with electron transfer dissociation capability peptide fragmentation notation. While cid remains the major fragmentation method for peptide sequencing, electron transfer dissociation etd is emerging as a complementary method for the characterization of peptides and post. Electrontransfer dissociation etd is a method of fragmenting multiplycharged gaseous macromolecules in a mass spectrometer between the stages of tandem mass spectrometry msms. Our present study provides evidence that neutral so3 losses can also occur in electron capture dissociation and electrontransfer dissociation experiments.
Electron transfer dissociation etd is a method of fragmenting multiplycharged gaseous macromolecules in a mass spectrometer between the stages of tandem mass spectrometry msms. In much earlier work, mcluckey and coworkers reported gasphase electron transfer reactions of multiply deprotonated nucleic acids with rare gas cations 4. Electron transfer dissociation mass spectrometry of peptides. Here we demonstrate the facile structural characterization of electron transfer dissociation generated peptide fragments by infrared ion spectroscopy using the tunable freeelectron laser felix, aiding the elucidation of the underlying dissociation mechanisms. A mass spectrometer is disclosed comprising an electron transfer dissociation device comprising an ion guide. Demonstrate electron transfer between atoms to form ionic bonds. Difference of electron capture and transfer dissociation mass. The inventive substances produce radical anions that lead to electron transfer dissociations with a high yield of fragment ions. Electron transfer dissociation mass spectrometry of. While cid remains the major fragmentation method for peptide sequencing, electron transfer dissociation etd is emerging as a. Introduction of freeradical sites to multiply charged peptides allows access to new fragmentation pathways that are otherwise too costly e.
Control of electron localization in molecular dissociation. Mass spectrometry has rapidly evolved to become the platform of choice for proteomic analysis. Electron transfer dissociation of peptide anions springerlink. Glycation is the nonenzymatic glycosylation of free amine groups on.
Electron transfer dissociation etd, a relatively new peptideprotein fragmentation. Electron transfer dissociation etd user guide 17 basic operation and maintenance 1 to mount and adjust the cooling plate to mount and adjust the cooling plate tools needed torx 10 torx 20 1 turn the heater device so that the nose the part to which the cooling plate attaches is in a line with the octopole preadjustment. Global proteomic profiling of phosphopeptides using electron. Singly charged anthracene anions transfer an electron to multiply protonated peptides in a radio frequency quadrupole linear ion trap qlt and induce fragmentation of the peptide backbone along pathways that are analogous to those observed in. Experimental data from ion mobility measurements and electron transfer dissociation were combined with extensive computational analysis of ion structures and dissociation energetics for glyleuglyglylys cations and cation radicals. The list of acronyms and abbreviations related to etd electron transfer dissociation. Request pdf electron transfer dissociation mass spectrometry of peptides containing free cysteine using group xii metals as a charge carrier electron transfer dissociation etd has been used. Applications include peptide sequencing 3, 4, identification of posttranslational modifications 5, 6, 7 and studies of noncovalent. Many of the ions that form have eight electrons in their valence shell. Jul 20, 2015 formation and reshuffling of disulfide bonds in bovine serum albumin demonstrated using tandem mass spectrometry with collisioninduced and electron transfer dissociation ine rombouts 1 bert lagrain 1. Feb, 2007 electron transfer dissociation etd is a recently introduced mass spectrometric technique that provides a more comprehensive coverage of peptide sequences and posttranslational modifications.
Etd is used extensively with polymers and biological molecules such as. Performance characteristics of electron transfer dissociation mass. Welcome to professor donald hunts tutorial on peptide sequence analysis by electron transfer dissociation etd mass spectrometry. The role of electron transfer dissociation in modern proteomics. Structural identification of electron transfer dissociation. Electron transfer dissociation of modified peptides and proteins volume. We also report that such neutral losses may be reduced by fragmenting peptidealkali metal adducts, such as sodiated or potassiated peptides. Progress in research is hindered by analytical limitations, especially in biological areas in which sensitivity and dynamic range are critical to success. Electron attachment step in electron capture dissociation. Peptide and protein quantification using itraq with electron. Et is a mechanistic description of a redox reaction, wherein the oxidation state of reactant and product changes. The activity of nadphdependent electron transfer from fnr to fd scheme 2 was measured using cytochrome c cyt c as an arbitrary electron acceptor as reported by onda 21 with modification in the reaction mixture of 50 mm trishcl ph 7.
Our present study provides evidence that neutral so3 losses can also occur in electron capture dissociation and electron transfer dissociation experiments. Etd can be used to activate carboniodine bonds in a laserfree experiment. Electron capture dissociation ecd and electron transfer dissociation etd in. Moreover, it provides an accurate description of the electronic structure changes and of the nuclear reorganization associated with the reaction. Formation and reshuffling of disulfide bonds in bovine serum albumin demonstrated using tandem mass spectrometry with collisioninduced and electrontransfer dissociation ine rombouts 1. Here we demonstrate the facile structural characterization of electron transfer dissociation generated peptide fragments by infrared ion spectroscopy using the tunable free electron laser felix, aiding the elucidation of the underlying dissociation mechanisms. Electron transfer article about electron transfer by the. Here, we evaluated the use of etd for a global phosphoproteome analysis. Electron transfer dissociation mass spectrometry of peptides containing free cysteine using group xii metals as a charge carrier b author. Electron transfer dissociation how is electron transfer. Electron transfer dissociation etd is a recently introduced mass spectrometric technique that provides a more comprehensive coverage of peptide sequences and posttranslational modifications. Electron transfer dissociation the scientist magazine.
Sequencing larger intact proteins 3070 kda with activated ion electron transfer dissociation. Electroncapture dissociation ecd is a method of fragmenting gasphase ions for structure elucidation of peptides and proteins in tandem mass spectrometry. Similar to electroncapture dissociation, etd induces fragmentation of large, multiplycharged cations by transferring electrons to them. Formation and reshuffling of disulfide bonds in bovine serum. Peptide backbone cleavage then occurs by hydrogen radical abstraction from a backbone amide n to facilitate cleavage of the adjacent cc bond, thereby producing a and xtype product ions. It requires multiplycharged gasphase cations z2 and therefore it is typically limited to an electron spray ion source. Structural identification of electron transfer dissociation products in mass spectrometry using infrared ion spectroscopy. The fragmentation chemistry of peptides containing intrachain disulfide bonds was investigated under electron transfer dissociation etd. Electron transfer dissociation mass spectrometry in proteomics. Ina sense, etd is the ionion analog of electron capture dissociation 3the reagent anion is simply a vehicle for electron delivery.
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