The role of copper in cysteine oxidation: study of intra- and inter-molecular reactions in mass spectrometry

Prudent, Michel; Girault, Hubert H.

doi:10.1039/b817061d

Cited by 55 publications

(51 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Such reactions on cysteine residues are not uncommon in proteins. [37] It was further suggested that, in consequence, the disulfide would negatively affect CDO activity by obstructing access of substrate to the active site. Ye and co-workers put this hypothesis to the test by designing CDO variants where C164 was replaced by either serine or alanine, both amino acids incapable of forming a disulfide with substrate cysteine.…”

Section: Discussionmentioning

confidence: 99%

Influence of cysteine 164 on active site structure in rat cysteine dioxygenase

et al. 2016

View full text Add to dashboard Cite

Cysteine dioxygenase (CDO) is a non-heme mononuclear iron enzyme with unique structural features, namely an intramolecular thioether crosslink between cysteine 93 and tyrosine 157, and a disulfide between substrate L-cysteine and cysteine 164 in the entrance channel to the active site. We investigated how these posttranslational modifications affect catalysis through a kinetic, crystallographic and computational study. The enzyme kinetics of a C164S variant are identical to WT indicating that disulfide formation at C164 does not significantly impair access to the active site at physiological pH. However, at high pH the cysteine-tyrosine crosslink formation is enhanced in C164S. This supports the view that disulfide formation at position 164 can limit access to the active site. The C164S yielded crystal structures of unusual clarity in both resting state and with cysteine bound. Both show that the iron in the cysteine bound complex is a mixture of penta-and hexa-coordinate with a water molecule taking up the final site (60% occupancy), which is where dioxygen is believed to coordinate during turnover. The serine also displays stronger hydrogen bond interactions to a water bound to the amine of the substrate cysteine. However, the interactions between cysteine and iron appear unchanged. DFT calculations support this and show that WT and C164S have similar binding energies for the water molecule in the final site. This variant therefore provides evidence that WT also exists in this equilibrium between penta-and hexa-coordinate forms and presence of the sixth ligand does not strongly affect dioxygen binding.

show abstract

Section: Discussionmentioning

confidence: 99%

Influence of cysteine 164 on active site structure in rat cysteine dioxygenase

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Because the single-cysteine constructs are not inhibited by copper, these data argue that in each case the two cysteines participate in a novel Cu 2ϩ -binding site and thus are presumably within ϳ4 Å of one another. An alternative possibility is that Cu 2ϩ binds and directly oxidizes the cysteine pair to a disulfide (20). In either case, the proximity of these cysteine residues provides strong support for the overall homology model of NKCC1, and the inhibitory nature of the interaction supports a transport role of TM10 movement relative to TM12.…”

Section: Inhibitory Cross-linking Of Tm10 -11/12mentioning

confidence: 98%

Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion Transport and Transporter Activation Revealed by Internal Cross-linking between Transmembrane Domains 10 and 11/12

Monette

Somasekharan

Forbush

2014

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Na-K-Cl cotransporters (NKCCs) are responsible for volume and chloride homeostasis and chloride transport and are targets of loop diuretic drugs. Results: Cross-links between transmembrane domains (TMs) 10 and 11/12 of NKCC1 are identified as inhibitory and stimulatory. Conclusion: Activation of NKCC1 involves movement of TM12 relative to TM10. Significance: This identifies movement of TM12 as a key step in the molecular mechanism of NKCC activation.

show abstract

“…This electrode was made either of platinum wire both for blank and metal salt experiments or of copper or zinc. [22][23][24] The dual-channel microsprayer (see the Tagging of Phosphopeptides Section) was used with reservoirs stuck on it or in a holder. [25,26] These devices were coupled to an ion trap mass spectrometer.…”

Section: Methodsmentioning

confidence: 99%

“…[22] Copper is known to catalyze the oxidation of cysteine residues and Cu + has a particular affinity for the thiol moiety. The complexation and the influences of copper ions electrogenerated from a copper electrode were tested on different cysteine-containing peptides.…”

Section: Functional Esi: Study Of Copper Complexesmentioning

confidence: 99%

About the Electrospray Ionization Source in Mass Spectrometry: Electrochemistry and On-chip Reactions

Prudent¹,

Méndez²,

Roussel³

et al. 2009

Chimia

Self Cite

View full text Add to dashboard Cite

The present work shows that the electrochemical properties of electrospray ionization (ESI) can be used to add functions to the process. As example, we show how the choice of the electrode material can be used to study interactions between metal ions and biomolecules in mass spectrometry (MS). In positive ionization MS, an electrospray device acts as anode, which implies oxidation reactions. Sacrificial electrodes (made of copper or zinc) are used to supply the electrospray current and to produce cations that are able to react on-line with compounds of interest. Thus, the interactions between copper ions and ligands or peptides were investigated by using a copper electrode. Another example is the in situ electrogeneration of a dinuclear zinc(II) complex for the mass tagging of phosphopeptides when working with a zinc electrode. In order to perform these reactions on the same microchip, a dual-channel microsprayer was used, where one channel was dedicated to the tag electrogeneration and the other to the infusion of a phosphopeptides solution. Finally, this dual-channel microsprayer was used to study complexation at liquid-liquid interfaces in biphasic ESI-MS, such as thioether crowns and lead ions or peptides and phospholipids complexes. These examples illustrate the use of electrochemistry and on-chip reactions in ESI-MS analysis.

show abstract

The role of copper in cysteine oxidation: study of intra- and inter-molecular reactions in mass spectrometry

Cited by 55 publications

References 47 publications

Influence of cysteine 164 on active site structure in rat cysteine dioxygenase

Influence of cysteine 164 on active site structure in rat cysteine dioxygenase

Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion Transport and Transporter Activation Revealed by Internal Cross-linking between Transmembrane Domains 10 and 11/12

About the Electrospray Ionization Source in Mass Spectrometry: Electrochemistry and On-chip Reactions

Contact Info

Product

Resources

About