The role of hydrophobic interactions in ankyrin–spectrin complex formation

Kolondra, Adam; Lenoir, Marc; Wolny, Marcin; Czogalla, Aleksander; Overduin, Michael; Sikorski, Aleksander F.; Grzybek, Michał

doi:10.1016/j.bbamem.2010.07.024

Cited by 11 publications

(24 citation statements)

References 25 publications

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“…In order to selectively monitor CTxB binding to its lipid receptor in pure Ld or Lo membrane only, we used synthetic liposomes consisting of either DOPC (Ld) or SM/Chol (Lo) and 0.1 mol% of either GM1 or bdGM1. These liposomes were then used as capture specimen in electrochemiluminescence ELISA assays to quantitatively evaluate CTxB binding (Kolondra et al, 2010; Lingwood et al, 2011). Equal GSL content in liposomes was validated by thin-layer chromatography (TLC) analysis (Figure S6).…”

Section: Resultsmentioning

confidence: 99%

Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

et al. 2017

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Driven by interactions between lipids and proteins, biological membranes display lateral heterogeneity that manifests itself in a mosaic of liquid-ordered (Lo) or raft, and liquid-disordered (Ld) or non-raft domains with a wide range of different properties and compositions. In giant plasma membrane vesicles and giant unilamellar vesicles, specific binding of Cholera Toxin (CTxB) to GM1 glycolipids is a commonly used strategy to label raft domains or Lo membrane environments. However, these studies often use acyl-chain labeled bodipy-GM1 (bdGM1), whose headgroup accessibility and membrane order or phase partitioning may differ from those of GM1, rendering the interpretation of CTxB binding data quite problematic. To unravel the molecular basis of CTxB binding to GM1 and bdGM1, we explored the partitioning and the headgroup presentation of these gangliosides in the Lo and Ld phases using atomistic molecular dynamics simulations complemented by CTxB binding experiments. The conformation of both GM1 and bdGM1 was shown to be largely similar in the Lo and Ld phases. However, bdGM1 showed reduction in receptor availability when reconstituted into synthetic bilayer mixtures, highlighting that membrane phase partitioning of the gangliosides plays a considerable role in CTxB binding. Our results suggest that the CTxB binding is predominately modulated by the partitioning of the receptor to an appropriate membrane phase. Further, given that the Lo and Ld partitioning of bdGM1 differs from those of GM1, usage of bdGM1 for studying GM1 behavior in cells can lead to invalid interpretation of experimental data.

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Section: Resultsmentioning

confidence: 99%

Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

et al. 2017

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“…The recorded data were analyzed using GraphPad Prism 5.0 software using one site-specific binding algorithm. To measure the interactions between ankBDn and its mutants with the spectrin-binding domain (ankSBD) of ankyrin a previously published protocol was used [34]. K D (equilibrium dissociation constant) and maximal binding capacity (B max ) values were calculated according to the equation B = B max * Free/(Free+K D )+nonspecific binding.…”

Section: Methodsmentioning

confidence: 99%

Key Amino Acid Residues of Ankyrin-Sensitive Phosphatidylethanolamine/Phosphatidylcholine-Lipid Binding Site of βI-Spectrin

et al. 2011

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It was shown previously that an ankyrin-sensitive, phosphatidylethanolamine/phosphatidylcholine (PE/PC) binding site maps to the N-terminal part of the ankyrin-binding domain of β-spectrin (ankBDn). Here we have identified the amino acid residues within this domain which are responsible for recognizing monolayers and bilayers composed of PE/PC mixtures. In vitro binding studies revealed that a quadruple mutant with substituted hydrophobic residues W1771, L1775, M1778 and W1779 not only failed to effectively bind PE/PC, but its residual PE/PC-binding activity was insensitive to inhibition with ankyrin. Structure prediction and analysis, supported by in vitro experiments, suggests that “opening” of the coiled-coil structure underlies the mechanism of this interaction. Experiments on red blood cells and HeLa cells supported the conclusions derived from the model and in vitro lipid-protein interaction results, and showed the potential physiological role of this binding. We postulate that direct interactions between spectrin ankBDn and PE-rich domains play an important role in stabilizing the structure of the spectrin-based membrane skeleton.

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“…Mutational analysis of the spectrin-binding domain of erythroid ankyrin (ZU5A domain) showed that F917S substitution completely abolished the formation of the spectrin-ankyrin complex [45]. It has been suggested that this residue is crucial for the appropriate orientation of the spectrin-binding site in erythroid ankyrin.…”

Section: Relationship Between Ankyrin-binding and Lipid-binding Activmentioning

confidence: 99%

“…The major isoform of erythroid ankyrin is composed of three distinct regions: an N-terminal domain known as a membrane domain containing 24 ankyrin repeats, a central region known as a spectrin-binding domain containing three subdomains (ZU5A, ZU5B and UPA), and a C-terminal regulatory domain with its subdomain called the "death domain" (DD). Only the first of the two ZU5 domains (ZU5A) of erythroid ankyrin is directly responsible for binding β-spectrin [43,[45][46][47]. Furthermore, the ZU5A-ZU5B-UPA-DD tandem of erythroid ankyrin seems to be crucial for auto-regulation of ankyrin functions [46].…”

Section: +mentioning

confidence: 99%

“…Interestingly, although F917Ank does not make a direct contact with spectrin, it is localized in a dense hydrophobic core (916SFMV919) and interacts with F913Ank, which is crucial for spectrin binding (Fig. 1) [45]. Studies carried out by Czogalla et al [112,128] based on the spin-label mobility and spin-spin distance measurements via electron paramagnetic resonance spectroscopy of the ankyrin-binding domain of erythroid β-spectrin preceded the reports on the atomic structure of this region by other three groups [42,47,[129][130][131].…”

Section: Relationship Between Ankyrin-binding and Lipid-binding Activmentioning

confidence: 99%

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Spectrin and phospholipids — the current picture of their fascinating interplay

Bogusławska

Machnicka

Hryniewicz-Jankowska

et al. 2014

Cellular and Molecular Biology Letters

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Abbreviations used: ABD -actin-binding domain; AE1 -anion exchanger 1; Ankankyrin; CH -calponin homology domain; GPC -glycophorin C; MPP1 -membrane palmitoylated protein 1; PC -phosphatidylcholine; PE -phosphatidylethanolamine; PHpleckstrin homology domain; PI -phosphatidylinositol; PI(4,5)P 2 -phosphatidylinositol-4,5-bisphosphate; PS -phosphatidylserine; SH3 -SRC homology 3 domain; UPA -Unc5-PIDD-ankyrin domain; ZU5 -domain present in ZO-1 and Unc5-like netrin receptors Abstract: The spectrin-based membrane skeleton is crucial for the mechanical stability and resilience of erythrocytes. It mainly contributes to membrane integrity, protein organization and trafficking. Two transmembrane protein macro-complexes that are linked together by spectrin tetramers play a crucial role in attaching the membrane skeleton to the cell membrane, but they are not exclusive. Considerable experimental data have shown that direct interactions between spectrin and membrane lipids are important for cell membrane cohesion. Spectrin is a multidomain, multifunctional protein with several distinctive structural regions, including lipid-binding sites within CH tandem domains, a PH domain, and triple helical segments, which are excellent examples of ligand specificity hidden in a regular repetitive structure, as recently shown for the ankyrin-sensitive lipid-binding domain of beta spectrin. In this review, we summarize the state of knowledge about interactions between spectrin and membrane lipids.

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The role of hydrophobic interactions in ankyrin–spectrin complex formation

Cited by 11 publications

References 25 publications

Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

Key Amino Acid Residues of Ankyrin-Sensitive Phosphatidylethanolamine/Phosphatidylcholine-Lipid Binding Site of βI-Spectrin

Spectrin and phospholipids — the current picture of their fascinating interplay

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