Inhibition of the catalytic activity of rhodanese by S-nitrosylation using nitric oxide donors

Kwiecień, Inga; Sokołowska, Maria; Luchter-Wasylewska, Ewa; Włodek, Lidia

doi:10.1016/s1357-2725(03)00005-0

Cited by 9 publications

(11 citation statements)

References 35 publications

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“…Prior to testing the effect of NO with respect to platelet adhesion, it was necessary to establish that NO has no effect on LDH activity. As a free radical, NO is known to react with various enzymes, which could possibly change their function and efficacy in catalysis 35, 36. Previous studies using the LDH assay to assess the biocompatibility of polymers involved evaluating only physically30, 32 or chemically29, 31 modified surfaces, not a material that releases a reactive species such as NO.…”

Section: Resultsmentioning

confidence: 99%

“…As a free radical, NO is known to react with various enzymes, which could possibly change their function and efficacy in catalysis. 35,36 Previous studies using the LDH assay to assess the biocompatibility of polymers involved evaluating only physically 30,32 or chemically 29,31 modified surfaces, not a material that releases a reactive species such as NO. Consequently, it was crucial to verify that any ''observed'' reduction in LDH activity derived from experiments using PRP was indeed the results of less adhered platelets rather than any inhibition of LDH function in the presence of continuous NO production.…”

Section: Ldh Assaymentioning

confidence: 99%

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In vitro platelet adhesion on polymeric surfaces with varying fluxes of continuous nitric oxide release

Zhou

Meyerhoff

2007

J Biomedical Materials Res

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Nitric oxide (NO) is released by endothelial cells that line the inner walls of healthy blood vessels at fluxes ranging from 0.5 x 10(-10) to 4.0 x 10(-10) mol cm(-2) min(-1), and this continuous NO release contributes to the extraordinary thromboresistance of the intact endothelium. To improve the biocompatibility of blood-contacting devices, a biomimetic approach to release/generate NO at polymer/blood interfaces has been pursued recently (with NO donors or NO generating catalysts doped within polymeric coatings) and this concept has been shown to be effective in preventing platelet adhesion/activation via several in vivo animal studies. However, there are no reports to date describing any quantitative in vitro assay to evaluate the blood compatibilities of such NO release/generating polymers with controlled NO fluxes. Such a methodology is desired to provide a preliminary assessment of any new NO-releasing material, in terms of the effectiveness of given NO fluxes and NO donor amounts on platelet activity before the more complex and costly in vivo testing is carried out. In this article, we report the use of a lactate dehydrogenase assay to study in vitro platelet adhesion on such NO-releasing polymer surfaces with varying NO fluxes. Reduced platelet adhesion was found to correlate with increasing NO fluxes. The highest NO flux tested, 7.05 (+/-0.25) x 10(-10) mol cm(-2) min(-1), effectively reduced platelet adhesion to nearly 20% of its original level (from 14.0 (+/-2.1) x 10(5) cells cm(-2) to 2.96 (+/-0.18) x 10(5) cells cm(-2)) compared to the control polymer coating without NO release capability.

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Section: Resultsmentioning

confidence: 99%

Section: Ldh Assaymentioning

confidence: 99%

In vitro platelet adhesion on polymeric surfaces with varying fluxes of continuous nitric oxide release

Zhou

Meyerhoff

2007

J Biomedical Materials Res

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“…Furthermore, the structure also revealed that the protein overexpressed in E. coli is partially both S-sulfhydrated and S-nitrosylated and that the S-sulfhydration is likely present in two conformations [12]. It has been known that S-nitrosylation and S-sulfhydration are mutually inhibitory processes, which may play important roles in H 2 S/NO signaling [13]. It has also been suspected that the ubiquity and frequent presence of the rhodanese domains in multidomain proteins implies physiological functions other than cell detoxification [14].…”

Section: Introductionmentioning

confidence: 99%

S-Sulfhydration of the Catalytic Cysteine in the Rhodanese Domain of YgaP is Complex Dynamic Process

Tzitzilonis

Kwiatkowski²,

Riek³

2016

Matters (Zür.)

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“…However, it has also been suspected that the ubiquity of the rhodanese domains and their frequent presence in multidomain proteins imply physiological functions other than cell detoxification [21]. The interplay between S-sulfhydration and S-nitrosylation, which are mutually inhibitory processes, is important in H 2 S/NO signaling [23]. Therefore, it is interesting that a rhodanese domain can be found in the membrane proteome of E. coli.…”

Section: Introductionmentioning

confidence: 99%

“…Although the physiological role of YgaP is unknown, the catalytic loop of its rhodanese domain, which includes Cys63, is essential for the sulfur transfer activity [21,22,24]. It has been shown that S-nitrosylation inactivates the function of rhodanese domains by nitrosylation of the catalytically active Cys [20,23]. Here, we investigated structural and dynamical changes affected by S-nitrosylation in vitro by both NMR spectroscopy and X-ray crystallography.…”

Section: Introductionmentioning

confidence: 99%

S-Nitrosylation Induces Structural and Dynamical Changes in a Rhodanese Family Protein

Tzitzilonis

Nakamura

Kwiatkowski

et al. 2016

Journal of Molecular Biology

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S-Nitrosylation is well established as an important post-translational regulator in protein function and signaling. However, relatively little is known about its structural and dynamical consequences. We have investigated the effects of S-nitrosylation on the rhodanese domain of the Escherichia coli integral membrane protein YgaP by NMR, X-ray crystallography, and mass spectrometry. The results show that the active cysteine in the rhodanese domain of YgaP is subjected to two competing modifications: S-nitrosylation and S-sulfhydration, which are naturally occurring in vivo. It has been observed that in addition to inhibition of the sulfur transfer activity, S-nitrosylation of the active site residue Cys63 causes an increase in slow motion and a displacement of helix 5 due to a weakening of the interaction between the active site and the helix dipole. These findings provide an example of how nitrosative stress can exert action at the atomic level.

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Inhibition of the catalytic activity of rhodanese by S-nitrosylation using nitric oxide donors

Cited by 9 publications

References 35 publications

In vitro platelet adhesion on polymeric surfaces with varying fluxes of continuous nitric oxide release

In vitro platelet adhesion on polymeric surfaces with varying fluxes of continuous nitric oxide release

S-Sulfhydration of the Catalytic Cysteine in the Rhodanese Domain of YgaP is Complex Dynamic Process

S-Nitrosylation Induces Structural and Dynamical Changes in a Rhodanese Family Protein

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