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newer applications including characterization of PTMs, non-tryptic peptides and intact proteins. (Kim Review) |
newer applications including characterization of PTMs, non-tryptic peptides and intact proteins. (Kim Review) |
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Performance Characteristics <ref>{{Cite journal |
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| last = Good |
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| first = David M. |
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| last2 = Wirtala |
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| first2 = Matthew |
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| last3 = McAlister |
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| first3 = Graeme C. |
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| last4 = Coon |
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| first4 = Joshua J. |
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| date = 2007-11-01 |
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| title = Performance Characteristics of Electron Transfer Dissociation Mass Spectrometry |
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| url = http://www.mcponline.org/content/6/11/1942 |
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| journal = Molecular & Cellular Proteomics |
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| language = en |
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| volume = 6 |
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| issue = 11 |
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| pages = 1942–1951 |
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| doi = 10.1074/mcp.M700073-MCP200 |
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| issn = 1535-9476 |
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| pmid = 17673454 |
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}}</ref> |
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Review <ref>{{Cite journal|last=Zhang|first=Zhaorui|last2=Wu|first2=Si|last3=Stenoien|first3=David L.|last4=Paša-Tolić|first4=Ljiljana|date=2014-01-01|title=High-Throughput Proteomics|url=http://dx.doi.org/10.1146/annurev-anchem-071213-020216|journal=Annual Review of Analytical Chemistry|volume=7|issue=1|pages=427–454|doi=10.1146/annurev-anchem-071213-020216|pmid=25014346}}</ref> |
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new article<ref>{{Cite journal|last=Nilsson|first=Jonas|date=2016-01-18|title=Liquid chromatography-tandem mass spectrometry-based fragmentation analysis of glycopeptides|url=http://link.springer.com/article/10.1007/s10719-016-9649-3|journal=Glycoconjugate Journal|language=en|pages=1–12|doi=10.1007/s10719-016-9649-3|issn=0282-0080}}</ref> |
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== See also == |
== See also == |
New Sandbox for Mass Spec Class
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This is a user sandbox of Kmcke14. You can use it for testing or practicing edits. |
Editing electron-transfer dissociation page for LSU Mass Spec class (Chem 4558) Spring 201
Electron-transfer dissociation (ETD) is a method of fragmenting multiply-charged gaseous macromolecules in a mass spectrometer between the stages of tandem mass spectrometry (MS/MS).[1] Similar to electron-capture dissociation, ETD induces fragmentation of large, multiply-charged cations by transferring electrons to them.[2] ETD is used extensively with polymers and biological molecules such as proteins and peptides for sequence analysis.[3] Transferring an electron causes peptide backbone cleavage into c- and z-ions while leaving labile post translational modifications (PTM) intact.[4] The method was developed by Donald F. Hunt, Joshua Coon, John E. P. Syka and Jarrod Marto at the University of Virginia.[5]
Electron-capture dissociation (ECD) was developed in 1998 to fragment large proteins for mass spectrometric analysis.[6] Because ECD requires a large amount of near-thermal electrons (<0.2eV), originally it was used exclusively with Fourier transform ion cyclotron resonance mass spectrometry (FTICR), the most expensive form of MS instrumentation.[7] Less costly options such as quadrupole time-of-flight (Q-TOF), quadrupole ion trap (QIT) and linear quadrupole ion trap (QLT) instruments used the more energy-intensive collision-induced dissociation method (CID), resulting in random fragmentation of peptides and proteins.[8] In 2004 Syka et al announced the creation of ETD, a dissociation method similar to ECD, but using a low-cost, widely available commercial spectrometer. The first ETD experiments were run on a QLT mass spectrometer with an electrospray ionization (ESI) source. [9]
Several steps are involved in electron transfer dissociation. Usually a protein mixture is first separated using high performance liquid chromatography (HPLC). Next multiply-protonated precursor molecules are generated by electrospray ionization and injected into the mass spectrometer. (Only molecules with a charge of 2+ or greater can be used in ETD.) In order for an electron to be transferred to the positive precursor molecules radical anions are generated and put into the ion trap with them. During the ion/ion reaction an electron is transferred to the positively-charged protein or peptide, causing fragmentation along the peptide backbone. Finally the resultant fragments are mass analyzed.[10]
In the original ETD experiments anthracene (C14H10) was was used to generate reactive radical anions through negative chemical ionization.[9] Several polycyclic aromatic hydrocarbon molecules have been used in subsequent experiments, with fluoranthene currently the preferred reagent.[11] Fluoranthene has only about 40% efficiency in electron transfer, however, so other molecules with low electron affinity are being sought. [10]
When the precursor cations (proteins or peptides) and radical anions are combined in the ion trap an electron is transferred to the mulitply-charged cation. This forms an unstable positive radical cation with one less positive charge and an odd electron. Fragmentation takes place along the peptide backbone at a N− Cα bond, resulting in c- and z-type fragment ions.[3]
Fragmentation caused by ETD allows more complete protein sequence information to be obtained from ETD spectra than from CID tandem mass spectrometry. Because many peptide backbone c- and z- type ions are detected, almost complete sequence coverage of many peptides can be discerned from ETD fragmentation spectra.[12] Sequences of 15-40 amino acids at both the N-terminus and the C-terminus of the protein can be read using mass-to-charge values for the singly and doubly charged ions. These sequences, together with the measured mass of the intact protein, can be compared to database entries for known proteins and to reveal post-translational modifications.[13]
Electron transfer dissociation takes place in an ion trap mass spectrometer with an electrospray ionization source.
newer applications including characterization of PTMs, non-tryptic peptides and intact proteins. (Kim Review)
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{{cite journal}}
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Category:Tandem mass spectrometry