A novel derivatization method for peptides to increase sensitivity and backbone fragmentation in liquid secondary‐ion mass spectra

Kiplinger, Jeffrey P.; Contillo, Leonard G.; Hendrick, W. Lee; Grodski, Alex

doi:10.1002/rcm.1290061207

Cited by 5 publications

(3 citation statements)

References 33 publications

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“…Also, there is potential for the charge to be spread over several sites if either an arginine or additional lysine residues are present in the peptide. Kiplinger et al (1992) used pentafluorobenzoyl fluoride to derivatize peptides at the N-terminus, as shown in Scheme 16. The derivatization was useful for analyses by LSIMS in the positive and negative modes.…”

Section: Side-chain Derivativesmentioning

confidence: 99%

Charge derivatization of peptides for analysis by mass spectrometry

Roth¹,

Huang²,

Sadagopan³

et al. 1998

Mass Spectrom. Rev.

133

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Section: Side-chain Derivativesmentioning

confidence: 99%

Charge derivatization of peptides for analysis by mass spectrometry

Roth¹,

Huang²,

Sadagopan³

et al. 1998

Mass Spectrom. Rev.

133

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“…Localization of a positive charge at a particular site on a peptide (13,14) also profoundly affects the nature of the fragment ions observed in high-collision energy tandem mass spectrometry (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26). A protonated Arg or Lys, or a cationic derivative located near the N terminus of a peptide, promotes the formation of fragment ions containing the N terminus (a, b, c ions; the nomenclature for peptide fragment ions has been discussed in ref.…”

mentioning

confidence: 99%

A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spectrometry

Keough

Youngquist

Lacey

1999

Proc. Natl. Acad. Sci. U.S.A.

225

262

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A method has been developed for de novo peptide sequencing using matrix-assisted laser desorption ionization mass spectrometry. This method will facilitate biological studies that require rapid determination of peptide or protein sequences, e.g., determination of posttranslational modifications, identification of active compounds isolated from combinatorial peptide libraries, and the selective identification of proteins as part of proteome studies. The method involves fast, one-step addition of a sulfonic acid group to the N terminus of tryptic peptides followed by acquisition of postsource decay (PSD) fragment ion spectra. The derivatives are designed to promote efficient charge site-initiated fragmentation of the backbone amide bonds and to selectively enhance the detection of a single fragment ion series that contains the C terminus of the molecule (y-ions). The overall method has been applied to pmol quantities of peptides. The resulting PSD fragment ion spectra often exhibit uninterrupted sequences of 20 or more amino acid residues. However, fragmentation efficiency decreases considerably at amide bonds on the C-terminal side of Pro. The spectra are simple enough that de novo sequence tagging is routine. The technique has been successfully applied to peptide mixtures, to high-mass peptides (up to 3,600 Da) and to the unambiguous identification of proteins isolated from two-dimensional gel electrophoresis. The PSD spectra of these derivatized peptides often allow far more selective protein sequence database searches than those obtained from the spectra of native peptides.Knowledge of protein sequences is fundamentally important for understanding many physiological processes at the molecular level (1). Tandem mass spectrometry has become an increasingly essential tool for protein and peptide sequencing because of its speed, sensitivity, and applicability to complex mixtures (2). Recently, postsource decay (PSD) matrix-assisted laser desorption ionization (MALDI) mass spectrometry was developed for high-sensitivity peptide sequencing applications (3-8). Subpicomole limits of analysis were reported as a result of the high yield of fragment ions and the high ion transmission inherent with time-of-flight mass spectrometry (5). Kaufmann et al. (5) also noted several problems associated with PSD MALDI sequencing of peptides, including the complexity of the resulting fragmentation patterns and the lack of computer algorithms capable of interpreting the complex spectra. The recent incorporation of delayed extraction (DE) (9, 10), a technique designed to improve precursor-ion mass resolution and mass-measurement accuracy, reduced the rate of PSD fragmentation by at least an order of magnitude. As a result, many low-intensity precursor ions obtained by DE MALDI do not produce enough PSD fragmentation to allow derivation of even short sequence tags (7).Positively charged derivatives have been used in desorption mass spectrometry for many years to improve sensitivity by enhancing ionization efficiencies (11, 12...

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“…Most approaches to facilitate manual sequencing are based on chemical derivatization to reduce the complexity of the MS/MS spectra by directing the sequence‐determining ions so that dominantly either b‐ or y‐ions18 are obtained 19. One of the early derivatization methods is to derivatize the N ‐terminal with pentafluorobenzoyl fluoride 20. This derivatization enhances the signal intensity in negative ion mode and, more interestingly, dramatically increases the fragmentation efficiency in positive ion mode.…”

Section: Introductionmentioning

confidence: 99%

On‐target sample preparation of 4‐sulfophenyl isothiocyanate‐derivatized peptides using AnchorChip Targets

2007

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De novo sequencing of tryptic peptides by post source decay (PSD) or collision induced dissociation (CID) analysis using MALDI TOF-TOF instruments is due to the easy interpretation facilitated by the introduction of N-terminal sulfonated derivatives. Recently, a stable and cheap reagent, 4-sulfophenyl isothiocyanate (SPITC), has been successfully used for N-terminal derivatization. Previously described methods have always used desalting and concentration by reverse-phase chromatography prior to mass spectrometric analysis. Here we present an on-target sample preparation method based on AnchorChip target technology. The method was optimized for reduction of by-products and sensitivity with SPITC-derivatized tryptic BSA peptides, and successfully applied to protein identification from silver-stained two-dimensional electrophoretic gels of fish liver extracts. The method is simple and sensitive and allowed protein identification based on de novo sequencing and BLAST search from species with limited sequence information.

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A novel derivatization method for peptides to increase sensitivity and backbone fragmentation in liquid secondary‐ion mass spectra

Cited by 5 publications

References 33 publications

Charge derivatization of peptides for analysis by mass spectrometry

Charge derivatization of peptides for analysis by mass spectrometry

A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spectrometry

On‐target sample preparation of 4‐sulfophenyl isothiocyanate‐derivatized peptides using AnchorChip Targets

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