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1.
Chamot-Rooke J van der Rest G Dalleu A Bay S Lemoine J 《Journal of the American Society for Mass Spectrometry》2007,18(8):1405-1413
Electron capture dissociation (ECD) has become an alternative method to collision-activated dissociation (CAD) to avoid gas-phase cleavage of post-translational modifications carried by side chains from the peptide backbone. Nonetheless, as illustrated herein by the study of O-glycosylated and O-phosphorylated peptides, the extent of ECD fragmentations may be insufficient to cover the entire peptide sequence and to localize accurately these modifications. The present work demonstrates that the derivatization of peptides at their N-terminus by a phosphonium group improves dramatically and systematically the sequence coverage deduced from the ECD spectrum for both O-glycosylated and O-phosphorylated peptides compared with their native counterparts. The exclusive presence of N-terminal fragments (c-type ions) in the ECD spectra of doubly charged molecular cations simplifies peptide sequence interpretation. Thus, the combination of ECD and fixed charge derivatization appears as an efficient analytical tool for the extensive sequencing of peptides bearing labile groups. 相似文献
2.
Electron capture dissociation (ECD) has recently been shown in some cases to produce abundant N-terminal b-ion peptide fragments. These product ions are usually only observed when activation occurs via vibrational excitation as in collision-induced dissociation (CID). Here, we show that occurrence of b-ions in the ECD spectra of synthetic peptides are correlated with low gas-phase basicity and that the observed b-ion fragments are N-terminal products. Furthermore, all ECD spectra containing b-ions also had abundant losses of hydrogen and ammonia from the charge-reduced species. 相似文献
3.
Piotr Stefanowicz Monika Kijewska Zbigniew Szewczuk 《Journal of mass spectrometry : JMS》2009,44(7):1047-1052
The electron capture dissociation (ECD) of peptide‐derived Amadori products has been successfully applied for their sequencing. In contrast to the collision induced dissociation (CID), based on the vibrational excitation of peptides, the ECD method does not produce ions formed by fragmentation of the hexose moiety, that facilitates interpretation of the obtained spectra. The fragmentation spectrum is dominated by cn and z·n ions, providing the sufficient information for sequencing of peptides and establishing the location of glycated Lys residues in the peptide chain. The ECD experiments were conducted on a series of synthetic peptides and unseparated digests of glycated ubiquitin. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
4.
Monika Kijewska Dorota Gszczyk Remigiusz Bchor Piotr Stefanowicz Zbigniew Szewczuk 《Molecules (Basel, Switzerland)》2021,26(22)
Peptide modification by a quaternary ammonium group containing a permanent positive charge is a promising method of increasing the ionization efficiency of the analyzed compounds, making ultra-sensitive detection even at the attomolar level possible. Charge-derivatized peptides may undergo both charge remote (ChR) and charge-directed (ChD) fragmentation. A series of model peptide conjugates derivatized with N,N,N-triethyloammonium (TEA), 1-azoniabicyclo[2.2.2]octane (ABCO), 2,4,6-triphenylopyridinium (TPP) and tris(2,4,6-trimetoxyphenylo)phosphonium (TMPP) groups were analyzed by their fragmentation pathways both in collision-induced dissociation (CID) and electron-capture dissociation (ECD) mode. The effect of the fixed-charge tag type and peptide sequence on the fragmentation pathways was investigated. We found that the aspartic acid effect plays a crucial role in the CID fragmentation of TPP and TEA peptide conjugates whereas it was not resolved for the peptides derivatized with the phosphonium group. ECD spectra are mostly dominated by cn ions. ECD fragmentation of TMPP-modified peptides results in the formation of intense fragments derived from this fixed-charge tag, which may serve as reporter ion. 相似文献
5.
Stensballe A Jensen ON Olsen JV Haselmann KF Zubarev RA 《Rapid communications in mass spectrometry : RCM》2000,14(19):1793-1800
Analysis of phosphotyrosine and phosphoserine containing peptides by nano-electrospray Fourier transform ion cyclotron resonance (FTICR) mass spectrometry established electron capture dissociation (ECD) as a viable method for phosphopeptide sequencing. In general, ECD spectra of synthetic and native phosphopeptides appeared less complex than conventional collision activated dissociation (CAD) mass spectra of these species. ECD of multiply protonated phosphopeptide ions generated mainly c- and z(.)-type peptide fragment ion series. No loss of water, phosphate groups or phosphoric acid from intact phosphopeptide ions nor from the c and z(.) fragment ion products was observed in the ECD spectra. ECD enabled complete or near-complete amino acid sequencing of phosphopeptides for the assignment of up to four phosphorylation sites in peptides in the mass range 1400 to 3500 Da. Nano-scale Fe(III)-affinity chromatography combined with nano-electrospray FTMS/ECD facilitated phosphopeptide analysis and amino acid sequencing from crude proteolytic peptide mixtures. 相似文献
6.
Guan Z 《Journal of the American Society for Mass Spectrometry》2002,13(12):1443-1447
Electron capture dissociation (ECD) has been demonstrated to be an effective fragmentation technique for characterizing the site and structure of the fatty acid modification in ghrelin, a 28-residue growth-hormone-releasing peptide that has an unusual ester-linked n-octanoyl (C8:0) modification at Ser-3. ECD cleaves 21 of 23 possible backbone amine bonds, with the product ions (c and z· ions) covering a greater amino acid sequence than those obtained by collisionally activated dissociation (CAD). Consistent with the ECD nonergodic mechanism, the ester-linked octanoyl group is retained on all backbone cleavage product ions, allowing for direct localization of this labile modification. In addition, ECD also induces the ester bond cleavage to cause the loss of octanoic acid from the ghrelin molecular ion; the elimination process is initiated by the capture of an electron at the protonated ester group, which is followed by the radical-site-initiated reaction known as -cleavage. The chemical composition of the attached fatty acid can be directly obtained from the accurate Fourier transform ion cyclotron resonance (FTICR) mass measurement of the ester bond cleavage product ions. 相似文献
7.
Charge States of y Ions in the Collision-Induced Dissociation of Doubly Charged Tryptic Peptide Ions
Bonds that break in collision-induced dissociation (CID) are often weakened by a nearby proton, which can, in principle, be
carried away by either of the product fragments. Since peptide backbone dissociation is commonly charge-directed, relative
intensities of charge states of product y- and b-ions depend on the final location of that proton. This study examines y-ion
charge distributions for dissociation of doubly charged peptide ions, using a large reference library of peptide ion fragmentation
generated from ion-trap CID of peptide ions from tryptic digests. Trends in relative intensities of y2+ and y1+ ions are examined as a function of bond cleavage position, peptide length (n), residues on either side of the bond and effects
of residues remote from the bond. It is found that yn-2/b2 dissociation is the most sensitive to adjacent amino acids, that y2+/y1+ steadily increase with increasing peptide length, that the N-terminal amino acid can have a major influence in all dissociations,
and in some cases other residues remote from the bond cleavage exert significant effects. Good correlation is found between
the values of y2+/y1+ for the peptide and the proton affinities of the amino acids present at the dissociating peptide bond. A few deviations from
this correlation are rationalized by specific effects of the amino acid residues. These correlations can be used to estimate
trends in y2+/y1+ ratios for peptide ions from amino acid proton affinities. 相似文献
8.
Alex G. Harrison Cagdas Tasoglu Talat Yalcin 《Journal of the American Society for Mass Spectrometry》2013,24(10):1565-1572
The fragmentation reactions of the MH+ ions of Leu-enkephalin amide and a variety of heptapeptide amides have been studied in detail as a function of collision energy using a QqToF beam type mass spectrometer. The initial fragmentation of the protonated amides involves primarily formation of bn ions, including significant loss of NH3 from the MH+ ions. Further fragmentation of these bn ions occurs following macrocyclization/ring opening leading in many cases to bn ions with permuted sequences and, thus, to formation of non-direct sequence ions. The importance of these non-direct sequence ions increases markedly with increasing collision energy, making peptide sequence determination difficult, if not impossible, at higher collision energies. Figure
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9.
Yin H Chacon A Porter NA Masterson DS 《Journal of the American Society for Mass Spectrometry》2007,18(5):807-816
Protein identification is routinely accomplished by peptide sequencing using mass spectrometry (MS) after enzymatic digestion.
Site-specific chemical modification may improve peptide ionization efficiency or sequence coverage in mass spectrometry. We
report herein that amino group of lysine residue in peptides can be selectively modified by reaction with a peroxycarbonate
and the resulting lysine peroxycarbamates undergo homolytic fragmentation under conditions of low-energy collision-induced
dissociation (CID) in electrospray ionization (ESI) and matrix-assisted laser desorption and ionization (MALDI) MS. Selective
modification of lysine residue in peptides by our strategy can induce specific peptide cleavage at or near the lysine site.
Studies using deuterated analogues of modified lysine indicate that fragmentation of the modified peptides involves apparent
free-radical processes that lead to peptide chain fragmentation and side-chain loss. The formation of a-, c-, or z-types of
ions in MS is reminiscent of the proposed free-radical mechanisms in low-energy electron capture dissociation (ECD) processes
that may have better sequence coverage than that of the conventional CID method. This site-specific cleavage of peptides by
free radical- promoted processes is feasible and such strategies may aid the protein sequencing analysis and have potential
applications in top-down proteomics. 相似文献
10.
Andrew W. Jones Peter J. Winn Helen J. Cooper 《Journal of the American Society for Mass Spectrometry》2012,23(12):2063-2074
The radical ion chemistry of a suite of S-nitrosopeptides has been investigated. Doubly and triply-protonated ions of peptides NYCGLPGEYWLGNDK, NYCGLPGEYWLGNDR, NYCGLPGERWLGNDR, NACGAPGEKWAGNDK, NYCGLPGEKYLGNDK, NYGLPGCEKWYGNDK and NYGLPGEKWYGCNDK were subjected to electron capture dissociation (ECD), and collision-induced dissociation (CID). The peptide sequences were selected such that the effect of the site of S-nitrosylation, the nature and position of the basic amino acid residues, and the nature of the other amino acid side chains, could be interrogated. The ECD mass spectra were dominated by a peak corresponding to loss of ?NO from the charge-reduced precursor, which can be explained by a modified Utah-Washington mechanism. Some backbone fragmentation in which the nitrosyl modification was preserved was also observed in the ECD of some peptides. Molecular dynamics simulations of peptide ion structure suggest that the ECD behavior was dependent on the surface accessibility of the protonated residue. CID of the S-nitrosylated peptides resulted in homolysis of the S?CN bond to form a long-lived radical with loss of ?NO. The radical peptide ions were isolated and subjected to ECD and CID. ECD of the radical peptide ions provided an interesting comparison to ECD of the unmodified peptides. The dominant process was electron capture without further dissociation (ECnoD). CID of the radical peptide ions resulted in cysteine, leucine, and asparagine side chain losses, and radical-induced backbone fragmentation at tryptophan, tyrosine, and asparagine residues, in addition to charge-directed backbone fragmentation. 相似文献
11.
Mihalca R Kleinnijenhuis AJ McDonnell LA Heck AJ Heeren RM 《Journal of the American Society for Mass Spectrometry》2004,15(12):1869-1873
Electron capture dissociation at 86 K of the linear peptide Substance P produced just two backbone fragments, whereas at room
temperature eight backbone fragments were formed. Similarly, with the cyclic peptide gramicidin S, just one backbone fragment
was formed at 86 K but five at room temperature. The observation that some backbone scissions are active and others inactive,
when all involve N-Cα cleavages and have a high rate constant, indicates that the more specific fragments at low temperatures reflects the reduced
conformation heterogeneity at low temperatures. This is supported by reduced or inactive hydrogen loss, a channel that has
previously been shown to be affected by conformation. The conclusion that the ECD fragments are a snapshot of the conformational
(intramolecular solvation shell) heterogeneity helps explain how the relative intensities of ECD fragments can be different
on different instrument and highlights the common theme in methodologies used to increase sequence coverage, namely an increase
in the conformational heterogeneity of the precursor ion population. 相似文献
12.
Shigeo Hayakawa Shinya Matsumoto Mami Hashimoto Kenichi Iwamoto Hirofumi Nagao Michisato Toyoda Yasushi Shigeri Michiko Tajiri Yoshinao Wada 《Journal of the American Society for Mass Spectrometry》2010,21(9):1482-1489
Post-translational modifications (PTMs) of proteins are important in the activation, localization, and regulation of protein
function in vivo. The usefulness of electron capture dissociation (ECD) and electron-transfer dissociation (ETD) in tandem
mass spectrometry (MS/MS) using low-energy (LE) trap type mass spectrometer is associated with no loss of a labile PTM group
regarding peptide and protein sequencing. The experimental results of high-energy (HE) collision induced dissociation (CID)
using the Xe and Cs targets and LE-ETD were compared for doubly-phosphorylated peptides TGFLT(p)EY(p)VATR (1). Although HE-CID using the Xe target did not provide information on the amino acid sequence, HE-CID using the Cs target
provided all the z-type ions without loss of the phosphate groups as a result of HE-ETD process, while LE-ETD using fluoranthene
anion gave only z-type ions from z5 to z11. The difference in the results of HE-CID between the Xe and Cs targets demonstrated that HE-ETD process with the Cs target
took place much more dominantly than collisional activation. The difference between HE-ETD using Cs targets and LE-ETD using
the anion demonstrated that mass discrimination was much weaker in the high-energy process. HE-ETD was also applied to three
other phosphopeptides YGGMHRQEX(p)VDC (2: X = S, 3: X = T, 4: X = Y). The HE-CID spectra of the doubly-protonated phosphopeptides
(= [M + 2H]2+) of 2, 3, and 4 using the Cs target showed a very similar feature that the c-type ions from c7 to c11 and the z-type ions from z7 to z11 were formed via N-Cα bond cleavage without a loss of the phosphate group. 相似文献
13.
T. Yu. Samgina K. A. Artemenko V. A. Gorshkov S. V. Ogourtsov R. A. Zubarev A. T. Lebedev 《Rapid communications in mass spectrometry : RCM》2009,23(9):1241-1248
A high‐performance liquid chromatography nano‐electrospray ionization Fourier transform mass spectrometry (HPLC/nanoESI‐FTMS) approach involving recording of collision‐activated dissociation (CAD) and electron‐capture dissociation (ECD) spectra of an intact sample and two its modifications after performic oxidation and reduction followed by carboxamidomethylation helps to establish peptide profiles in the crude secretion of frog species at mid‐throughput level, including de novo sequencing. The proposed derivatization procedures allow increasing of the general sequence coverage in the backbone, providing complementary information and, what is more important, reveal the amino acid sequence in the cystine ring (‘rana box’). Thus purely mass spectrometric efficient sequencing becomes possible for longer than usual proteolytic peptides. Seventeen peptides belonging to four known families were identified in the secretion of the European brown frog Rana arvalis inhabiting the Moscow region in Russia. Ranatuerins, considered previously a unique feature of the North American species, as well as a new melittin‐related peptide, are worth special mention. The developed approach was previously successfully used for the identification of peptides in the skin secretion of the Caucasian green frog Rana ridibunda. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
14.
Huilin Li Jonathon R. Snelling Mark P. Barrow James H. Scrivens Peter J. Sadler Peter B. O’Connor 《Journal of the American Society for Mass Spectrometry》2014,25(7):1217-1227
To further explore the binding chemistry of cisplatin (cis-Pt(NH3)2Cl2) to peptides and also establish mass spectrometry (MS) strategies to quickly assign the platinum-binding sites, a series of peptides with potential cisplatin binding sites (Met(S), His(N), Cys(S), disulfide, carboxyl groups of Asp and Glu, and amine groups of Arg and Lys, were reacted with cisplatin, then analyzed by electron capture dissociation (ECD) in a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS). Radical-mediated side-chain losses from the charge-reduced Pt-binding species (such as CH3S? or CH3SH from Met, SH? from Cys, CO2 from Glu or Asp, and NH2 ? from amine groups) were found to be characteristic indicators for rapid and unambiguous localization of the Pt-binding sites to certain amino acid residues. The method was then successfully applied to interpret the top-down ECD spectrum of an inter-chain Pt-crosslinked insulin dimer, insulin?+?Pt(NH3)2?+?insulin (>10 kDa). In addition, ion mobility MS shows that Pt binds to multiple sites in Substance P, generating multiple conformers, which can be partially localized by collisionally activated dissociation (CAD). Platinum(II) (Pt(II)) was found to coordinate to amine groups of Arg and Lys, but not to disulfide bonds under the conditions used. The coordination of Pt to Arg or Lys appears to arise from the migration of Pt(II) from Met(S) as shown by monitoring the reaction products at different pH values by ECD. No direct binding of cisplatin to amine groups was observed at pH 3?~?10 unless Met residues were present in the sequence, but noncovalent interactions between cisplatin hydrolysis and amination [Pt(NH3)4]2+ products and these peptides were found regardless of pH. Figure
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15.
Zhao C Xie B Chan SY Costello CE O'Connor PB 《Journal of the American Society for Mass Spectrometry》2008,19(1):138-150
Doubly charged sodiated and permethylated linear malto-oligosaccharides ({Glc}6-{Glc}9), branched N-linked glycans (high-mannose type GlcNAc2Man5-9, and complex asialo- and disialylated-biantennary glycans) were analyzed by tandem mass spectrometry using collisionally-activated dissociation (CAD) and "hot" electron capture dissociation (ECD) available in a custom-built ESI FTICR mass spectrometer. For linear permethylated malto-oligosaccharides, both CAD and "hot" ECD produced glycosidic cleavages (B, Y, C, and Z ions), cross-ring cleavages (A- and X-type), and internal cleavages (B/Y and C/Y type) to provide sequence and linkage information. For the branched N-linked glycans, CAD and "hot" ECD provided complementary structural information. CAD generated abundant B and Y fragment ions by glycosidic cleavages, whereas "hot" ECD produced dominant C and Z ions. A-type cross-ring cleavages were present in CAD spectra. Complementary A- and X-type cross-ring fragmentation pairs were generated by "hot" ECD, and these delineated the branching patterns and linkage positions. For example, 0, 4An and 3, 5An ions defined the linkage position of the major branch as the 6-position of the central core mannose residue. The internal fragments observed in CAD were more numerous and abundant than in "hot" ECD spectra. Since the triply charged (sodiated) molecular ion of the permethylated disialylated-biantennary N-linked glycan has relatively high abundance, it was isolated and fragmented in a "hot" ECD experiment and extensive fragment ions (glycosidic and complementary pairs of cross-ring cleavages) were generated to fully confirm the sequence, branching, and linkage assignments for this glycan. 相似文献
16.
Xiaojuan Li Cheng Lin Liang Han Catherine E. Costello Peter B. O’Connor 《Journal of the American Society for Mass Spectrometry》2010,21(4):646-656
Secondary fragmentations of three synthetic peptides (human αA crystallin peptide 1-11, the deamidated form of human βB2 crystallin peptide 4-14, and amyloid β peptide 25-35) were studied in both electron capture dissociation (ECD) and electron-transfer dissociation (ETD) mode. In
ECD, in addition to c and z· ion formations, charge remote fragmentations (CRF) of z· ions were abundant, resulting in internal
fragment formation or partial/entire side-chain losses from amino acids, sometimes several residues away from the backbone
cleavage site, and to some extent multiple side-chain losses. The internal fragments were observed in peptides with basic
residues located in the middle of the sequences, which was different from most tryptic peptides with basic residues located
at the C-terminus. These secondary cleavages were initiated by hydrogen abstraction at the α-, β-, or γ-position of the amino acid side chain. In comparison, ETD generates fewer CRF fragments than ECD. This secondary cleavage
study will facilitate ECD/ETD spectra interpretation, and help de novo sequencing and database searching. 相似文献
17.
Hisham Ben Hamidane Aleksey Vorobyev Maud Larregola Aneta Lukaszuk Dirk Tourwé Dr. Solange Lavielle Philippe Karoyan Dr. Yury O. Tsybin Dr. 《Chemistry (Weinheim an der Bergstrasse, Germany)》2010,16(15):4612-4622
We report on the characteristics of the radical‐ion‐driven dissociation of a diverse array of β‐amino acids incorporated into α‐peptides, as probed by tandem electron‐capture and electron‐transfer dissociation (ECD/ETD) mass spectrometry. The reported results demonstrate a stronger ECD/ETD dependence on the nature of the amino acid side chain for β‐amino acids than for their α‐form counterparts. In particular, only aromatic (e.g., β‐Phe), and to a substantially lower extent, carbonyl‐containing (e.g., β‐Glu and β‐Gln) amino acid side chains, lead to N? Cβ bond cleavage in the corresponding β‐amino acids. We conclude that radical stabilization must be provided by the side chain to enable the radical‐driven fragmentation from the nearby backbone carbonyl carbon to proceed. In contrast with the cleavage of backbones derived from α‐amino acids, ECD of peptides composed mainly of β‐amino acids reveals a shift in cleavage priority from the N? Cβ to the Cα? C bond. The incorporation of CH2 groups into the peptide backbone may thus drastically influence the backbone charge solvation preference. The characteristics of radical‐driven β‐amino acid dissociation described herein are of particular importance to methods development, applications in peptide sequencing, and peptide and protein modification (e.g., deamidation and isomerization) analysis in life science research. 相似文献
18.
Malgorzata A. Kaczorowska Anna C. G. Hotze Michael J. Hannon Helen J. Cooper 《Journal of the American Society for Mass Spectrometry》2010,21(2):300-309
The electron capture dissociation (ECD) of metallo-supramolecular dinuclear triple-stranded helicate Fe2L34+ ions was determined by Fourier transform ion cyclotron resonance mass spectrometry. Initial electron capture by the di-iron(II)
triple helicate ions produces dinuclear double-stranded complexes analogous to those seen in solution with the monocationic
metal centers CuI or AgI. The gas-phase fragmentation behavior [ECD, collision-induced dissociation (CID), and infrared multiphoton dissociation (IRMPD)]
of the di-iron double-stranded complexes, (i.e., MS3 of the ECD product) was compared with the ECD, CID, and IRMPD of the CuI and AgI complexes generated from solution. The results suggest that iron-bound dimers may be of the form Fe2IL22+ and that ECD by metallo-complexes allows access, in the gas phase, to oxidation states and coordination chemistry that cannot
be accessed in solution. 相似文献
19.
Olsen JV Haselmann KF Nielsen ML Budnik BA Nielsen PE Zubarev RA 《Rapid communications in mass spectrometry : RCM》2001,15(12):969-974
Electron capture dissociation (ECD) in Fourier transform ion cyclotron resonance mass spectrometry coupled with electrospray ionization enhances the sequence elucidation of peptide nucleic acids compared with conventional low-energy collisionally activated dissociation (CAD). Examples are shown where ECD produced complete or extensive sequence coverage in PNAs six to ten nucleobases long. However, facile base losses from the reduced species and low abundances of backbone ECD fragments presented a significant problem. This was rationalized through the lower degree of charge solvation on the backbone compared to polypeptides. Combination of both CAD and ECD data is advantageous, as these techniques produce cleavages at different sites. 相似文献
20.
Anastasia Kalli Sonja Hess 《Journal of the American Society for Mass Spectrometry》2012,23(10):1729-1740
Hydrogen-deficient peptide radical cations exhibit fascinating gas phase chemistry, which is governed by radical driven dissociation and, in many cases, by a combination of radical and charge driven fragmentation. Here we examine electron capture dissociation (ECD) of doubly, [M + H]2+?, and triply, [M + 2H]3+?, charged hydrogen-deficient species, aiming to investigate the effect of a hydrogen-deficient radical site on the ECD outcome and characterize the dissociation pathways of hydrogen-deficient species in ECD. ECD of [M + H]2+? and [M + 2H]3+? precursor ions resulted in efficient electron capture by the hydrogen-deficient species. However, the intensities of c- and z-type product ions were reduced, compared with those observed for the even electron species, indicating suppression of N?CC?? backbone bond cleavages. We postulate that radical recombination occurs after the initial electron capture event leading to a stable even electron intermediate, which does not trigger N?CC?? bond dissociations. Although the intensities of c- and z-type product ions were reduced, the number of backbone bond cleavages remained largely unaffected between the ECD spectra of the even electron and hydrogen-deficient species. We hypothesize that a small ion population exist as a biradical, which can trigger N?CC?? bond cleavages. Alternatively, radical recombination and N?CC?? bond cleavages can be in competition, with radical recombination being the dominant pathway and N?CC?? cleavages occurring to a lesser degree. Formation of b- and y-type ions observed for two of the hydrogen-deficient peptides examined is also discussed. 相似文献