Na /K -ATPase interaction with methylglyoxal as reactive metabolic side product
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F17%3A73582007" target="_blank" >RIV/61989592:15110/17:73582007 - isvavai.cz</a>
Alternative codes found
RIV/00209805:_____/17:00077887
Result on the web
<a href="https://ac.els-cdn.com/S0891584917301739/1-s2.0-S0891584917301739-main.pdf?_tid=37409eb4-c2e3-11e7-a507-00000aacb362&acdnat=1509966860_7109c31ebd4d3a12da2c5260fc2e404b" target="_blank" >https://ac.els-cdn.com/S0891584917301739/1-s2.0-S0891584917301739-main.pdf?_tid=37409eb4-c2e3-11e7-a507-00000aacb362&acdnat=1509966860_7109c31ebd4d3a12da2c5260fc2e404b</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.freeradbiomed.2017.03.024" target="_blank" >10.1016/j.freeradbiomed.2017.03.024</a>
Alternative languages
Result language
angličtina
Original language name
Na /K -ATPase interaction with methylglyoxal as reactive metabolic side product
Original language description
Proteins are subject to oxidative modification and the formation of adducts with a broad spectrum of reactive species via enzymatic and non-enzymatic mechanisms. Here we report that in vitro non-enzymatic methylglyoxal (MGO) binding causes the inhibition and formation of MGO advanced glycation end-products (MAGEs) in Na+/K+-ATPase (NKA). Concretely, MGO adducts with NKA amino acid residues (mainly Arg) and Nε-(carboxymethyl)lysine (CML) formation were found. MGO is not only an inhibitor for solubilized NKA (IC50=91±16μM), but also for reconstituted NKA in the lipid bilayer environment, which was clearly demonstrated using a DPPC/DPPE liposome model in the presence or absence of the NKA-selective inhibitor ouabain. High-resolution mass spectrometric analysis of a tryptic digest of NKA isolated from pig (Sus scrofa) kidney indicates that the intracellular α-subunit is naturally (post-translationally) modified by MGO in vivo. In contrast to this, the β-subunit could only be modified by MGO artificially, and the transmembrane part of the protein did not undergo MGO binding under the experimental setup used. As with bovine serum albumin, serving as the water-soluble model, we also demonstrated a high binding capacity of MGO to water-poorly soluble NKA using a multi-spectral methodology based on electroanalytical, immunochemical and fluorimetric tools. In addition, a partial suppression of the MGO-mediated inhibitory effect could be observed in the presence of aminoguanidine (pimagedine), a glycation suppressor and MGO-scavenger. All the results here were obtained with the X-ray structure of NKA in the E1 conformation (3WGV) and could be used in the further interpretation of the functionality of this key enzyme in the presence of highly-reactive metabolic side-products, glycation agents and generally under oxidative stress conditions.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Free Radical Biology & Medicine
ISSN
0891-5849
e-ISSN
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Volume of the periodical
108
Issue of the periodical within the volume
July
Country of publishing house
US - UNITED STATES
Number of pages
9
Pages from-to
146-154
UT code for WoS article
000403463500014
EID of the result in the Scopus database
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