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Title: Structural Determination of Lysine-Linked Cisplatin Complexes via IRMPD Action Spectroscopy: NN s and NO – Binding Modes of Lysine to Platinum Coexist
Award ID(s):
1709789 1357887 0922819
NSF-PAR ID:
10383307
Author(s) / Creator(s):
; ; ; ; ; ; ; ; ; ;
Date Published:
Journal Name:
The Journal of Physical Chemistry B
Volume:
126
Issue:
45
ISSN:
1520-6106
Page Range / eLocation ID:
9246 to 9260
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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  1. Rationale

    N6‐Formyl lysine is a well‐known modification of histones and other proteins. It can also be formed as a damaged product from direct formylation of free lysine and accompanied by other lysine derivatives such as acetylated or methylated forms. In relation to the activity of cellular repair enzymes in protein turnover and to lysine metabolism, it is important to accurately quantify the overall ratio of modified lysine to free lysine.

    Methods

    N6‐Formyl lysine was quantified using liquid chromatography/tandem mass spectrometry (LC/MS/MS) with data collected in a non‐targeted manner using positive mode electrospray ionization on a Q‐Exactive HF+Orbitrap mass spectrometer. Studies were performed with lysine and deuterated lysine spiked into protein digests and solvents to investigate the extent of spontaneous formation and matrix effects of formation ofN6‐formyl lysine.

    Results

    We show thatN6‐formyl lysine,N2‐formyl lysine,N6‐acetyl lysine, andN2‐acetyl lysine are all formed spontaneously during sample preparation and LC/MS/MS analysis, which complicates quantification of these metabolites in biological samples.N6‐Formyl lysine was spontaneously formed and correlated to the concentration of lysine. In the sample matrix of protein digests, 0.03% of lysine was spontaneously converted intoN6‐formyl lysine, and 0.005% of lysine was converted intoN6‐formyl lysine in pure run solvent.

    Conclusions

    Spontaneous formation ofN6‐formyl lysine,N6‐acetyl lysine,N2‐formyl lysine, andN2‐acetyl lysine needs to be subtracted from biologically formed lysine modifications when quantifying these epimetabolites in biological samples.

     
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