Fragmentation patterns of chromophore‐tagged peptides in visible laser induced dissociation

Abstract

LID of derivatized cysteine-containing peptides displays the initial fragmentation of the chromophore. As energy is redistributed all along the peptide sequence, fragmentation of the peptide backbone is also observed. Thus, LID of chromophore-tagged peptides produces adequate fragment ions, allowing both good sequence coverage for a greater confidence of identification, and a large choice of transitions for specific quantification.

Data-Independent Acquisition Coupled to Visible Laser-Induced Dissociation at 473 nm (DIA-LID) for Peptide-Centric Specific Analysis of Cysteine-Containing Peptide Subset

“…The spectrum is dominated by the intense reporter ion at m / z 252.113. This could be explained by preferential protonation on the chromophore moiety which drives the fragmentation mechanism in HCD (charge-driven fragmentation), while the energetic redistribution seems more efficient in LID . As for backbone fragments, due to intense contributions from other cysteine-free ions in the DIA window, only a 2 + , b 2 + , and b 3 + fragment ions can be clearly assigned to the aforementioned kinase related peptide.…”

Increasing specificity of tandem mass spectrometry by laser‐induced dissociation

“…After the initial absorption at the chromophore site, energy is redistributed all along the peptide backbone. The fragmentation efficiency principally depends on the peptide length and the location of the modification . The LID spectra of NN‐, Alexa Fluor 350‐ and Dabcyl‐derivatized peptides also present a reporter ion arising from internal fragmentation of the chromophores.…”

“…The fragmentation efficiency principally depends on the peptide length and the location of the modification. 42 The LID spectra of NN-, Alexa Unfortunately, when radical-directed dissociation at 266 nm 28,29,36,37 is used, the formation of a radical species induces fragmentation only in the vicinity of the initial radical in subsequent CID, which is valuable to easily identify the site of modification but does (Figure 7). Other labelling could be envisaged but it should be kept in mind that complex sample preparation could be deleterious for quantitative analysis due to incomplete reaction yields and risks of losing compounds.…”

Unbiased Detection of Cysteine Sulfenic Acid by 473 nm Photodissociation Mass Spectrometry: Toward Facile In Vivo Oxidative Status of Plasma Proteins

“…With the aim of targeting Cys-containing peptide, this concept has been implemented using the judicious selection of chromophores absorbing in the UV range at 351 nm 30 and 266 nm 31,32 or in the visible range at 473 nm. [33][34][35] In the present work, we unveil a strategy coupling mass spectrometry and LID at 473 nm for streamlining the detection of cysteine sulfenic acid-containing peptides in complex protein digests. A new Dabcyl chromophore probe functionalized with a cyclohexanedione group was designed in order to ensure controlled grafting of SOH moieties, hence conferring specific absorption property at 473 nm of oxidized peptides.…”

“…The added value of the exquisite fragmentation specificity provided by laser-induced dissociation (LID) to detect a peptide subpopulation has been highlighted many times, for instance, with infrared multiphoton photodissociation at 10.6 μm for targeting sulfonated peptides. , Intrinsic chromophores such as disulfide bonds or tyrosine residues have been similarly used for specific detection in the ultraviolet range (UVPD). , The grafting of amino-acid side chains with a proper chromophore is an alternative for introducing specificity at the fragmentation step. With the aim of targeting a Cys-containing peptide, this concept has been implemented using the judicious selection of chromophores absorbing in the UV range at 351 and 266 nm , or in the visible range at 473 nm. − …”