Tuning of the Na,K-ATPase by the beta subunit

Hilbers, Florian; Kopeć, Wojciech; Isaksen, Toke Jost; Holm, Thomas Hellesøe; Lykke‐Hartmann, Karin; Nissen, Poul; Khandelia, Himanshu; Poulsen, Hanne

doi:10.1038/srep20442

Cited by 41 publications

(48 citation statements)

References 63 publications

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“…This effect of beta on the pump was found to be due to the TM helix rather than the N- or C-terminal domains, and specifically to a difference between the three isoforms in the tilt angles of their TM helices (Hilbers et al, 2016). …”

Section: Betasmentioning

confidence: 99%

The Structure and Function of the Na,K-ATPase Isoforms in Health and Disease

2017

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The sodium and potassium gradients across the plasma membrane are used by animal cells for numerous processes, and the range of demands requires that the responsible ion pump, the Na,K-ATPase, can be fine-tuned to the different cellular needs. Therefore, several isoforms are expressed of each of the three subunits that make a Na,K-ATPase, the alpha, beta and FXYD subunits. This review summarizes the various roles and expression patterns of the Na,K-ATPase subunit isoforms and maps the sequence variations to compare the differences structurally. Mutations in the Na,K-ATPase genes encoding alpha subunit isoforms have severe physiological consequences, causing very distinct, often neurological diseases. The differences in the pathophysiological effects of mutations further underline how the kinetic parameters, regulation and proteomic interactions of the Na,K-ATPase isoforms are optimized for the individual cellular needs.

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Section: Betasmentioning

confidence: 99%

The Structure and Function of the Na,K-ATPase Isoforms in Health and Disease

2017

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“…55 Moreover, sites N118 and N238 have been shown to be important in mammalians systems for folding and localization of the Na/K-ATPase complex to the plasma membrane, 55 where it creates concentration gradients important for a variety of cell physiological functions. The tilt of the transmembrane helix of the β2 subunit, which is close to N118, N153, and N238 glycosites, mediates functional differences in the Na/K-ATPase complex, 56 suggesting that various glycans at these sites also have the potential to alter function via conformational changes. The β1 subunit of Na/K-ATPase is also glycosylated (all three known sites are also characterized in this dataset), but the importance of specific glycosites is less pronounced in β1 versus β2 subunits, 57,58 highlighting the differential roles glycosylation heterogeneity can play even within isoforms of the same complex.…”

Section: Visualizing Glycoproteome Heterogeneitymentioning

confidence: 99%

Capturing site-specific heterogeneity with large-scale N-glycoproteome analysis

Riley

Hebert

Westphall

et al. 2019

Preprint

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Protein glycosylation is a highly important, yet a poorly understood protein post-translational modification. Thousands of possible glycan structures and compositions create potential for tremendous site heterogeneity and analytical challenge. A lack of suitable analytical methods for large-scale analyses of intact glycopeptides has ultimately limited our abilities to both address the degree of heterogeneity across the glycoproteome and to understand how it contributes biologically to complex systems. Here we show that N-glycoproteome site-specific microheterogeneity can be captured via large-scale glycopeptide profiling with methods enabled by activated ion electron transfer dissociation (AI-ETD), ultimately characterizing 1,545 Nglycosites (>5,600 unique N-glycopeptides) from mouse brain tissue. Moreover, we have used this large-scale glycoproteomic data to develop several new visualizations that will prove useful for analyzing intact glycopeptides in future studies. Our data reveal that N-glycosylation profiles can differ between subcellular regions and structural domains and that N-glycosite heterogeneity manifests in several different forms, including dramatic differences in glycosites on the same protein. MAIN TEXTAs insights into the role of glycosylation in health and disease continue to emerge, new technologies are required to improve the depth and breadth of glycoproteome analysis. [1][2][3][4] Especially needed are methods for intact glycopeptide analysisan approach that preserves biological context of the modification and enables understanding of proteome wide glycan heterogeneity. 5,6 The recent investment in glycoscience technology development made by the National Institutes of Health Common Fund program evince this importance and need. 7 Mass spectrometry (MS)-based methods are the premier approach to glycoproteome characterization.The bulk of our glycoproteome knowledge comes from methods that enzymatically cleave the glycan from the peptide and then sequence each molecular class separately. This approach has revealed a stunning diversity of hundreds of unique N-linked glycan structures that can decorate proteins. The specific residues that carried the modification can likewise be identified; however, the information of which glycan structures go on which sites is lost. For this reason, one cannot determine whether particular sites or classes of proteins have preference for one structure or another nor ultimately what the functional roles of the various glycans are. It is known that glycan heterogeneity can affect structure and function, such as binding specificities in immunoglobulins, 8,9 but the functional effects of heterogeneity across the glycoproteome remain largely uncharacterized.To gain a better understanding of glycan heterogeneity at a given site one can analyze intact glycopeptides. Similar to many other post-translational modifications (PTMs), glycopeptides require enrichment prior to analysis because of low stoichiometry and suppressed ionization efficiency compared to unmodified p...

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“…We also found the b3 isoform to be unchanged with gestation. Although not extensively studied in intact preparations, this isoform is thought to influence ion transport by modifying ionic affinities (Jaisser et al 1994;Hilbers et al 2016). Our data would point to these being essential properties for the Na + , K + ATPase expressed in myometrium at all stages of pregnancy.…”

Section: Discussionmentioning

confidence: 76%

“…; Hilbers et al. ). Our data would point to these being essential properties for the Na + , K + ATPase expressed in myometrium at all stages of pregnancy.…”

Section: Discussionmentioning

confidence: 97%

Gestation changes sodium pump isoform expression, leading to changes in ouabain sensitivity, contractility, and intracellular calcium in rat uterus

2017

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Developmental and tissue‐specific differences in isoforms allow Na+, K+‐ATPase function to be tightly regulated, as they control sensitivity to ions and inhibitors. Uterine contraction relies on the activity of the Na+, K+ ATPase, which creates ionic gradients that drive excitation‐contraction coupling. It is unknown whether Na+, K+ ATPase isoforms are regulated throughout pregnancy or whether they have a direct role in modulating uterine contractility. We hypothesized that gestation‐dependent differential expression of isoforms would affect contractile responses to Na+, K+ ATPase α subunit inhibition with ouabain. Our aims were therefore: (1) to determine the gestation‐dependent expression of mRNA transcripts, protein abundance and tissue distribution of Na+, K+ ATPase isoforms in myometrium; (2) to investigate the functional effects of differential isoform expression via ouabain sensitivity; and (3) if changes in contractile responses can be explained by changes in intracellular [Ca2+]. Changes in abundance and distribution of the Na+, K+ ATPase α, β and FXYD1 and 2 isoforms, were studied in rat uterus from nonpregnant, and early, mid‐, and term gestation. All α, β subunit isoforms (1,2,3) and FXYD1 were detected but FXYD2 was absent. The α1 and β1 isoforms were unchanged throughout pregnancy, whereas α2 and α3 significant decreased at term while β2 and FXYD1 significantly increased from mid‐term onwards. These changes in expression correlated with increased functional sensitivity to ouabain, and parallel changes in intracellular Ca2+, measured with Indo‐1. In conclusion, gestation induces specific regulatory changes in expression of Na+, K+ ATPase isoforms in the uterus which influence contractility and may be related to the physiological requirements for successful pregnancy and delivery.

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Tuning of the Na,K-ATPase by the beta subunit

Cited by 41 publications

References 63 publications

The Structure and Function of the Na,K-ATPase Isoforms in Health and Disease

The Structure and Function of the Na,K-ATPase Isoforms in Health and Disease

Capturing site-specific heterogeneity with large-scale N-glycoproteome analysis

Gestation changes sodium pump isoform expression, leading to changes in ouabain sensitivity, contractility, and intracellular calcium in rat uterus

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