The thermal unfolding and domain structure of Na<sup>+</sup>/K<sup>+</sup>‐exchanging ATPase.

Grinberg, Asya V.; Gevondyan, N. M.; Grinberg, Natalia V.; Grinberg, V. Ya.

doi:10.1046/j.0014-2956.2001.02436.x

Cited by 36 publications

(35 citation statements)

References 59 publications

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“…Based on the calorimetric data, the thermodynamic analysis of the protein folding allows the elucidation of proteins tertiary structure features and determination of interaction contributions that maintain the stability of the native structure [21].…”

Section: Resultsmentioning

confidence: 99%

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Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Yoneda

Rigos

Lourenço

et al. 2014

Archives of Biochemistry and Biophysics

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Differential scanning calorimetry (DSC) was applied to investigate the effect of cholesterol on the thermotropic properties of the lipid membrane (DPPC and DPPE). The thermostability and unfolding of solubilized and reconstituted Na,K-ATPase in DPPC:DPPE:cholesterol-liposomes was also studied to gain insight into the role of cholesterol in the Na,K-ATPase modulation of enzyme function and activity. The tertiary system (DPPC:DPPE:cholesterol) (molar ratio DPPC:DPPE equal 1:1) when cholesterol content was increased from 0% up to 40% results in a slight decrease in the temperature of transition and enthalpy, and an increase in width. We observed that, without heating treatment, at 37°C, the activity was higher for 20mol% cholesterol. However, thermal inactivation experiments showed that the enzyme activity loss time depends on the cholesterol membrane content. The unfolding of the enzyme incorporated to liposomes of DPPC:DPPE (1:1mol) with different cholesterol contents, ranging from 0% to 40% mol was also studied by DSC. Some differences between the thermograms indicate that the presence of lipids promotes a conformational change in protein structure and this change is enough to change the way Na,K-ATPase thermally unfolds.

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Section: Resultsmentioning

confidence: 99%

“…Differential scanning calorimetry (DSC) 1 is known to be one of the most powerful methods for studying protein unfolding, as it provides information on the structural organization and interactions of cooperative domains in proteins [21].…”

Section: Introductionmentioning

confidence: 99%

Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Yoneda

Rigos

Lourenço

et al. 2014

Archives of Biochemistry and Biophysics

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“…Normally, the purified renal Na,K-ATPase (␥-containing) is stable at 37°C, and although with differential scanning microcalorimetry, the heat capacity profile begins at that temperature (59). Most interestingly, the related Ca 2ϩ -ATPase SERCA1a, which does not have a subunit corresponding to ␤, begins to denature at 37°C (60).…”

Section: Discussionmentioning

confidence: 99%

Na,K-ATPase from Mice Lacking the γ Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability

Jones

Arystarkhova

et al. 2005

Journal of Biological Chemistry

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The ␥ subunit of the Na,K-ATPase, a 7-kDa single-span membrane protein, is a member of the FXYD gene family. Several FXYD proteins have been shown to bind to Na,K-ATPase and modulate its properties, and each FXYD protein appears to alter enzyme kinetics differently. Different results have sometimes been obtained with different experimental systems, however. To test for effects of ␥ in a native tissue environment, mice lacking a functional ␥ subunit gene (Fxyd2) were generated. These mice were viable and without observable pathology. Prior work in the mouse embryo showed that ␥ is expressed at the blastocyst stage. However, there was no delay in blastocele formation, and the expected Mendelian ratios of offspring were obtained even with Fxyd2 ؊/؊ dams. In adult Fxyd2 ؊/؊ mouse kidney, splice variants of ␥ that have different nephron segment-specific expression patterns were absent. Purified ␥-deficient renal Na,K-ATPase displayed higher apparent affinity for Na ؉ without significant change in apparent affinity for K ؉ . Affinity for ATP, which was expected to be decreased, was instead slightly increased. The results suggest that regulation of Na ؉ sensitivity is a major functional role for this protein, whereas regulation of ATP affinity may be context-specific. Most importantly, this implies that ␥ and other FXYD proteins have their effects by local and not global conformation change. Na,K-ATPase lacking the ␥ subunit had increased thermal lability. Combined with other evidence that ␥ participates in an early step of thermal denaturation, this indicates that FXYD proteins may play an important structural role in the enzyme complex.

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“…Formation of a loose-bundle state is consistent with results obtained with other techniques from other membrane proteins, which also indicate a large retention of secondary structure during the denaturation of the proteins. [47][48][49][50] It will be interesting to see how this loose-bundle state relates to the intermediate state postulated in the two-stage models for the folding of ␣-helical membrane proteins.…”

Section: Denaturation Of Rhodopsin Leads To Formation Of Loose-bundlementioning

confidence: 99%

Fourier transform IR spectroscopy study for new insights into molecular properties and activation mechanisms of visual pigment rhodopsin

Vogel

Siebert

2003

Biopolymers

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Fourier transform IR (FTIR) spectroscopy has been successfully applied in recent years to examine the functional and structural properties of the membrane protein rhodopsin, a prototype G protein coupled receptor. Unlike UV-visible spectroscopy, FTIR spectroscopy is structurally sensitive. It may give us both global information about the conformation of the protein and very detailed information about the retinal chromophore and all other functional groups, even when these are not directly related to the chromophore. Furthermore, it can be successfully applied to the photointermediates of rhodopsin, including the active receptor species, metarhodopsin II, and its decay products, which is not expected presently or even in the near future from crystallographic approaches. In this review we show how FTIR spectroscopy has significantly contributed to the understanding of very different aspects of rhodopsin, comprising both structural properties and the mechanisms leading to receptor activation and deactivation.

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The thermal unfolding and domain structure of Na⁺/K⁺‐exchanging ATPase.

Cited by 36 publications

References 59 publications

Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Na,K-ATPase from Mice Lacking the γ Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability

Fourier transform IR spectroscopy study for new insights into molecular properties and activation mechanisms of visual pigment rhodopsin

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The thermal unfolding and domain structure of Na+/K+‐exchanging ATPase.

Cited by 36 publications

References 59 publications

Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Na,K-ATPase reconstituted in ternary liposome: The presence of cholesterol affects protein activity and thermal stability

Na,K-ATPase from Mice Lacking the γ Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability

Fourier transform IR spectroscopy study for new insights into molecular properties and activation mechanisms of visual pigment rhodopsin

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The thermal unfolding and domain structure of Na⁺/K⁺‐exchanging ATPase.