Electron transfer dissociation etd based topdown mass spectrometry ms is the method of choice for indepth characterization of posttranslationally modified large peptides, small and mediumsized proteins, and noncovalent protein complexes. Electron transfer dissociation of amide nitrogen methylated polypeptide cations, journal of the american society for mass spectrometry, 2011, pp. Hence, the nonergodic fragmentation propensity of electron transfer dissociation etd fragmentation. A common use of tandemms is the analysis of biomolecules, such as proteins and peptides the molecules of a given sample are ionized and the first.
The cid spectrum shows a few b and y ions, but is dominated by a peak corresponding to the loss of phosphoric acid. In etd, the electron originates from a molecular anion and transfer results in charge reduction, energy increase, and dissociation. Etd is a valuable tandem ms method for characterizing many aspects of the proteome and is used in bottomup, middledown, and topdown proteomic methods. Negative electron transfer dissociation fourier transform. What hinders electron transfer dissociation etd of dna. Electron transfer dissociation etd of multiprotonated 6 20mer oligonucleotides and 12 and 14mer duplexes is compared to collision activated dissociation cad.
Jan 28, 2020 adpribosylation adpr is a posttranslational modification that plays pivotal roles in a wide range of cellular processes. Electrontransfer dissociation etd is a method of fragmenting multiplycharged gaseous. Electron transfer dissociation mass spectrometry in. 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 etd user guide 3 in this guide this guide provides information about your etd source. Structural characterization of glycosaminoglycans gags has been a challenge in the field of mass spectrometry, and the application of electron detachment dissociation edd fourier transform ion cyclotron resonance mass spectrometry fticrms has shown great promise to gag oligosaccharide characterization in a single tandem mass spectrometry experiment.
Electron transfer dissociation is a supremely useful tool for the analysis of posttranslationally modified proteins. 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. The luminal helix l of psab is essential for recognition of plastocyanin or cytochrome c 6 and fast electron transfer to photosystem i in chlamydomonas reinhardtii. Negative electron transfer dissociation of glycosaminoglycans. Structural analysis of intact monoclonal antibodies by. Rationale the modes of cleavage of lanthioninemethyllanthionine bridges under electron transfer dissociation etd were investigated using. Here we describe the first implementation of aietd on a quadrupoleorbitrapquadrupole linear ion trap qlt hybrid ms system orbitrap fusion lumos and demonstrate the. Analysis of intact monoclonal antibody igg1 by electron transfer. Electron transfer dissociation etd, utilizes radical anions produced in a chemical ionization ci source external to the trap. Review open access middledown electron capture dissociation and electron transfer dissociation for histone analysis annie moradian1, catarina franco1,2, michael j. Electron attachment step in electron capture dissociation ecd and electron transfer dissociation etd iwona anusiewicz, joanna berdyskochanska, and jack simons chemistry department and henry eyring center for theoretical chemistry, university of utah, salt lake city, utah 84112 received. Mechanical properties of 3d nanostructures obtained by.
Orbitrap fourier transform mass spectrometer ftms, to analyze a commercial. Electron attachment step in electron capture dissociation. Results are presented for the dissociative ionization of deuterium molecules d 2. Kayhooi khoo, in comprehensive natural products ii, 2010. Etd is also better suited for investigating phosphorylation and glycosylation. Electron capture dissociation ecd and electron transfer dissociation etd in metalpeptide complexes are dependent on the metal cation in. Electron transfer dissociation etd has been developed recently as an efficient ion fragmentation technique in mass spectrometry ms, being presently considered a step forward in proteomics with real perspectives for improvement, upgrade and application. Structural analysis of intact monoclonal antibodies by electron transfer dissociation mass spectrometry. It is one of the most widely used techniques for activation and dissociation of mass selected precursor ion in msms. A limitation of etd is the requirement that analytes be at least doubly cationized by. Fundamentals of electron capture dissociation fourier. The inventive substances produce radical anions that lead to electron transfer dissociations with a high yield of fragment ions. Electron transfer dissociation of oligonucleotide cations. Electroncapture dissociation an overview sciencedirect.
Sulfopeptide fragmentation in electroncapture and electrontransfer dissociation. The secondorder rate constants k 2 and the dissociation constants k d determined for wild. Peptide and protein sequence analysis by electron transfer. Electron shuttling in electron transfer dissociation. Electron transfer dissociation etd is an important tandem mass spectrometry technique in peptide and protein sequencing. Etd capability can be added to many thermo scientific ion. 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. Electron transfer dissociation of peptide anions article in journal of the american society for mass spectrometry 166. Middledown electron capture dissociation and electron.
To apply electron transfer dissociation etd mass spectrometry to topdown protein sequencing to apply and optimize such an approach for protein n and cterminal sequencing experimental sample preparation standard proteins were purchased from sigma. Electrontransfer dissociation etd is a variation of ecd, in which an ionion. For convenience in this discussion we will assume d and a are neutral molecules so that electrostatic forces may be ignored. Using concurrent ir photoactivation during electron transfer dissociation etd reactions, i. The role of electron transfer dissociation in modern proteomics. 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. Electron transfer dissociation of synthetic and natural peptides. Sulfopeptide fragmentation in electron capture and electron transfer dissociation journal of the american society for mass spectrometry, sep 2007 k.
Electron transfer dissociation allows differentiation of isoaspartic acid and aspartic acid residues using the same c 57 and z 57 peaks that were previously observed with electron capture dissociation. Both the radical anions for electron transfer dissociation and the nonradical anions for deprotonation are produced in a single ion source from. Etd causes efficient charge reduction of the multiprotonated oligonucleotides in addition to limited backbone cleavages to yield sequence ions of low abundance. Control of electron localization in molecular dissociation. Much excitement has been generated by the advent of electron capture dissociation ecd 144,145 and more recently the electron transfer dissociation etd 146,147 modes of msms, as applied to glycopeptides. Electron transfer dissociation etd bruker daltonics. Implementation of efficient electron transfer dissociation. Electron capture dissociation ecd and electron transfer dissociation etd of doubly protonated electron affinity eatuned peptides were studied to further illuminate the mechanism of these processes. However, peptide identification rates from etd spectra compare poorly with those from collision induced dissociation cid spectra, especially for doubly charged precursors. Identification of human hemoglobin protein variants using. A mass spectrometer is disclosed comprising an electron transfer dissociation device comprising an ion guide. Trastuzumab is known to have one nlinked glycosylation site on the.
Analytical and computational studies of intramolecular. Woski yonghyun kim a dissertation submitted in partial fulfillment of the requirements for the degree of doctor of philosophy in the department of chemistry. 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. Mass spectrometry has rapidly evolved to become the platform of choice for proteomic analysis. Electron transfer dissociation etd bruker daltonics pdf. The substances have high volatility that allows them to be kept in. The low energy, radicaldriven fragmentation techniques electron capture dissociation ecd and electron transfer dissociation etd are also employed to avoid excess vibrational energy. Electron transfer dissociation etd is a powerful new tool for peptide structure analysis and protein identi.
Electrontransfer dissociation an overview sciencedirect. Here, we describe the applicability of activated ion electron transfer dissociation aietd for. Available also on affordable ion trap mass spectrometers, etd induces specific nc. 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. In contrast, the etd spectrum displays a rich fragmentation pattern of c and z ions that reveals the exact location of the phosphorylation t. May 01, 2010 to facilitate electron ion reactions in quadrupole ion trap mass spectrometers, ionion reactions were developed to induce electron transfer. Charge state dependent topdown characterisation using. In mass spectrometric analysis equipment, highly charged analyte fragment ions are obtained by electron transfer dissociation of highly charged analyte ions by radical anions and the dissociation products are then deprotonated with nonradical anions. Difference of electron capture and transfer dissociation mass. Electron transfer dissociation etdbased topdown mass spectrometry ms is the method of choice for indepth characterization of posttranslationally modified large peptides, small and mediumsized proteins, and noncovalent protein complexes. Both are electron driven dissociation techniques and are. Pdf structural analysis of intact monoclonal antibodies. Sulfopeptide fragmentation in electroncapture and electron. Electrontransfer dissociation an overview sciencedirect topics.
Analytical and computational studies of intramolecular electron transfer pertinent to electron transfer and electron capture dissociation mass spectrometry diane neff and jack simons chemistry department and henry eyring center for theoretical chemistry university of utah, salt lake city, utah 84112. To achieve single amino acid resolution, we examined the extents of hdx by using electron transfer dissociation etd fragmentation of peptides representing selected regions of both the monomeric and the oligomeric forms of apoe4. From these experiments, we could identify the specific residues involved in apoe oligomerization. Electron transfer dissociation of amide nitrogen methylated. Electron transfer dissociation of nlinked glycopeptides. Electrontransfer dissociation etd is a method of fragmenting multiplycharged gaseous macromolecules in a mass spectrometer between the stages of tandem mass spectrometry msms. Electron shuttling in electron transfer dissociation article in international journal of mass spectrometry 2831.
Us20110062323a1 electron transfer dissociation device. Marcus theory of electron transfer from a molecular perspective, marcus theory is typically applied to outer sphere et between an electron donor d and an electron acceptor a. It requires multiplycharged gasphase cations z2 and therefore it is typically limited to an electron spray ion source. Electron transfer dissociation is a popular peptide fragmentation technique in mass spectrometry. Analytical and computational studies of intramolecular electron transfer pertinent to electron transfer and electron capture dissociation mass spectrometry diane neff and jack simons chemistry department and henry eyring center for theoretical chemistry university of. Jun 29, 2004 peptide sequence analysis using a combination of gasphase ionion chemistry and tandem mass spectrometry msms is demonstrated. Recent developments and applications of electron transfer.
Part spectra from 30minute tryptic digests of a normal hb top and the variant containing hb bottom. Adpribosylation adpr is a posttranslational modification that plays pivotal roles in a wide range of cellular processes. Etd capability can be added to many thermo scientific ion trap and orbitrap mass spectrometers. The luminal helix l of psab is essential for recognition. Here, we applied electron transfer dissociation etd, implemented on a hybrid. Electron transfer dissociation is a radicaldriven fragmentation technique which has found increasing use in the structural analysis of peptides and proteins. Both are electrondriven dissociation techniques and are. Partially purified daf16 protein fragment was a gift from nibs, beijing, china. The mechanisms of electron capture and electron transfer dissociation ecd and etd are investigated by covalently attaching a freeradical hydrogen atom. Charge state dependent topdown characterisation using electron.
Herein, electron transfer dissociation of 3fold protonated dna hexamers was studied to gain insight into the fragmentation mechanism. Ina sense, etd is the ionion analog of electron capture dissociation 3the reagent anion is simply a vehicle for electron delivery. Sep 20, 2017 radical activation methods, such as electron transfer dissociation etd, produce structural information complementary to collisioninduced dissociation. In this method, multiply protonated peptides or proteins are confined in the penning trap of. In the past, etd experiments have primarily involved basic peptides. Chapter 1 basic operation and maintenance this chapter describes basic operation and maintenance of the etd source. Identification of human hemoglobin protein variants using electrospray ionization electron transfer dissociation mass spectrometry 3. Structural identification of electron transfer dissociation products in. Similar to electroncapture dissociation, etd induces fragmentation of large, multiplycharged cations by transferring electrons to them. 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 recent years, electron transfer dissociation etd has enjoyed widespread applications from sequencing of peptides with or without posttranslational modifications to topdown analysis of intact proteins. Automated hydrogendeuterium exchange electron transfer.
Implementation of efficient electron transfer dissociation on. These peaks clearly define both the presence and the position of isoaspartic acid residues and they are relatively abundant. Sequencing by the increasingly popular electroninduced dissociation methods exd, such as electron capture and transfer dissociation, ecd. Electron transfer dissociation mass spectromerty studies of peptides by changgeng feng carolyn j. Improved peptide identification for proteomic analysis. Similar to electron capture dissociation, etd induces fragmentation of large, multiplycharged cations by transferring electrons to them. Electron transfer dissociation etd utilizes radical anions produced in a chemical ionization ci source external to the trap. Product specifications electron transfer dissociation etd. Bruker daltonics leader in etd technology amazon featuring etd since its commercial introduction in 2005, bruker has been the leader in highly efficient and sensitive electron transfer dissociation etd and proton transfer reaction ptr technology. Peptide sequence analysis using a combination of gasphase ionion chemistry and tandem mass spectrometry msms is demonstrated. Electron transfer dissociation is a radicaldriven fragmentation technique which has found increasing use in the structural an\. Herein, a novel dissociation pathway is reported, yielding homolytic cleavage of carboniodine bonds via electronic excitation.
Dissociative electron transfer electrochemical society. 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. We demonstrated how the subcycle evolution of the electric field of light can be used to control the motion of bound electrons. Development and application of novel electron transfer. Here, we describe the applicability of activated ion electron transfer dissociation aietd.
Here we report the first demonstration of nearcomplete sequence coverage of intact proteins using activated ion electron transfer dissociation aietd, a method that leverages concurrent infrared photoactivation to enhance electron driven dissociation. Electron transfer dissociation mass spectrometry of. Electron capture dissociation ecd is a method of fragmenting gasphase ions for structure elucidation of peptides and proteins in tandem mass spectrometry. A mass spectrometrybased assay combining the specificity of selected reaction monitoring and the protein ion activation capabilities of electron transfer dissociation was developed and employed for the rapid identification of hemoglobin variants from whole blood without previous proteolytic cleavage. Electron transfer dissociation 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. Electrontransfer dissociation allows differentiation of isoaspartic acid and aspartic acid residues using the same c 57 and z 57 peaks that were previously observed with electron capture dissociation. The sample, usually in solution, is first ionized by electrospray. Leveraging electron transfer dissociation for site selective radical. Electron transfer dissociation of peptide anions request pdf. Sweredoski1 and sonja hess1 abstract the posttranslational modifications ptms of histones play a major role in activating or silencing gene transcription. Tandem mass spectrometry, also known as msms or ms 2, is a technique in instrumental analysis where two or more mass analyzers are coupled together using an additional reaction step to increase their abilities to analyse chemical samples. Aug 28, 2015 electron transfer dissociation is a popular peptide fragmentation technique in mass spectrometry. Electron transfer dissociation etd data suggested optimal sequence coverage for. Electron transfer dissociation of nlinked glycopeptides from a recombinant mab using synapt g2s hdms 3 tryptic recombinant monoclonal igg, trastuzumab, was used for the etd analysis.
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