IndraLab

Statements

CALM1 binds HP. 12 / 12
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"The resulting list of HP-CAM peptides was further filtered to include peptides of a select list of potential protein targets."
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"An important caveat to the MRM method, however, is that method development is more time-consuming, and in the case of APAP-modified peptides for this study, the custom alkylation HP-CAM peptides were crucial for confirmation."
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"Results from scheduled MRM experiments, including peak areas for each transition monitored, relative areas, and retention times and calculated deviations to the control HP-CAM peptides ( xref )."
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"HP-CAM modification yields a positional isomer of APAP-modification on all cysteines and modified peptides were identified using the same criteria as APAP-modified peptides from database searching."
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"The mouse peptide specifically was a very long peptide with 33 residues and a charge state of 4+, making it quite difficult to screen for via LC-MRM without a corresponding signal in the HP-CAM reference sample."
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"HP-CAM peptides were used to confirm which fractions to monitor for each modified peptide, as well as LC retention time and MS/MS fragmentation behavior, with relatively high signal intensity."
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"In total, 1466 HP-CAM-modified proteins with 4554 distinct HP-CAM peptides were identified in the mouse control sample; and 1073 HP-CAM-modified proteins with 3599 distinct HP-CAM-peptides were found in the rat control sample."
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"APAP-treated and HP-CAM alkylated liver digests were analyzed by scheduled LC-MRM methods specific to each SPE fraction."
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"This large peptide was not easily amenable to MRM analysis due to the lack of significantly intense fragment ions, as well as no appropriate HP-CAM peptide for method optimisation."
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"MRM transitions of modified peptides were also monitored in adjacent fractions as those where they were detected in HP-CAM samples, since peptides can often elute in more than one fraction."
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"As for sorbitol dehydrogenase, the corresponding ortholog peptide was not incorporated into the MRM method, since it was not found in the rat HP-CAM reference sample and thus could not be optimized in the targeted method."
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"LC-MRM peaks were integrated, and relative peak areas for each transition (transition/sum of all transitions) as well as retention times were compared with HP-CAM signals for the same peptide. xref shows a representative example of a confirmed target LC-MRM for a mouse CYP 2C29 peptide, found via data-dependent MS/MS and confirmed by MRM analyses."
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