2002
DOI: 10.1021/pr025518b
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Defining Absolute Confidence Limits in the Identification of Caulobacter Proteins by Peptide Mass Mapping

Abstract: A derivatization reaction, guanidination, was recently reported that increases MALDI-TOF MS sensitivity toward lysine-terminated peptides. Its application conveys sequence information that can be used as a parameter in peptide mass mapping database searches. This paper presents a systematic study of the impact of guanidination on proteomic analysis of an entire bacterial organelle. Sixty-two 2-D gel isolated proteins from Caulobacter crescentus stalks were studied. A novel computer algorithm, Prodigies, was de… Show more

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Cited by 25 publications
(35 citation statements)
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“…Spot analysis was performed as described previously (Karty et al , 2002b). Briefly, spots were cut from the second‐dimension gels using cut‐off P200 pipette tips.…”
Section: Methodsmentioning
confidence: 99%
“…Spot analysis was performed as described previously (Karty et al , 2002b). Briefly, spots were cut from the second‐dimension gels using cut‐off P200 pipette tips.…”
Section: Methodsmentioning
confidence: 99%
“…The identification of the N-terminal residue provides a database searching constraint. We simulated the application of this constraint using the translated genome of Caulobacter crescentus and a database analysis program (PRODIGIES) that was written in house [34,35]. When candidate sequences are limited by both the mass of a precursor ion and the identity of the N-terminal residue, the number of candidate sequences is reduced by approximately one order of magnitude.…”
Section: Fragmentation Of Electrosprayed Amidine Labeled Peptidesmentioning
confidence: 99%
“…This phenomenon led to the development of signal-amplified tags (Cagney and Emili, 2002;Karty et al, 2002;Reilly, 2002, 2003). A mass-coded abundance tagging (MCAT) method was reported which involved the derivatization of the ε-amino groups of Lys residues using Omethylisourea; this labeling process causes a 42-Da mass difference (Cagney and Emili, 2002), as indicated in Fig.…”
Section: Guanidination and Gunidination-like Labelingmentioning
confidence: 99%
“…The guanidination (or guanidination-like modification) does indeed improve the ionization of peptides and the signals of y ions, which could be used for de novo sequencing. In fact, 30% more proteins were identified from the Caulobacter proteome after guanidination (Karty et al, 2002). However, this MCAT (or QUEST) method possessed two major drawbacks for the large-scale analysis of protein expression.…”
Section: Guanidination and Gunidination-like Labelingmentioning
confidence: 99%