Calpain‐induced proteolysis of β‐spectrins

Löfvenberg, Lars; Backman, Lars

doi:10.1016/s0014-5793(98)01697-4

Cited by 32 publications

(22 citation statements)

References 31 publications

(37 reference statements)

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“…3, one can see that the increased Ca 2+ content also leads to a vesiculation of the cells, in agreement with findings reported in literature [31,32]. Calpain, a calcium-dependent proteolytic enzyme, cleaves spectrin and actin, leading to cytoskeleton breakdown [40][41][42], and therefore may be involved in vesiculation.…”

Section: Discussionsupporting

confidence: 85%

Regulation of Phosphatidylserine Exposure in Red Blood Cells

Nguyen

Wagner-Britz

Maia

et al. 2011

Cell Physiol Biochem

113

121

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The exposure of phosphatidylserine (PS) on the outer membrane leaflet of red blood cells (RBCs) serves as a signal for eryptosis, a mechanism for the RBC clearance from blood circulation. The process of PS exposure was investigated as function of the intracellular Ca²⁺ content and the activation of PKCα in human and sheep RBCs. Cells were treated with lysophosphatidic acid (LPA), 4-bromo-A23187, or phorbol-12 myristate-13 acetate (PMA) and analysed by flow cytometry, single cell fluorescence video imaging, or confocal microscopy. For human RBCs, no clear correlation existed between the number of cells with an elevated Ca²⁺ content and PS exposure. Results are explained by three different mechanisms responsible for the PS exposure in human RBCs: (i) Ca²⁺-stimulated scramblase activation (and flippase inhibition) by LPA, 4-bromo-A23187, and PMA; (ii) PKC activation by LPA and PMA; and (iii) enhanced lipid flop caused by LPA. In sheep RBCs, only the latter mechanism occurs suggesting absence of scramblase activity.

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Section: Discussionsupporting

confidence: 85%

Regulation of Phosphatidylserine Exposure in Red Blood Cells

Nguyen

Wagner-Britz

Maia

et al. 2011

Cell Physiol Biochem

113

121

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“…Other cleavages of βII spectrin by calpain also exist (52 , 53), yet beyond the identification in fusion peptides of a susceptible site near the COOH terminus (A 2067 ) of βII spectrin (53), the precise locus of any calpain-cleavage site in βII spectrin or its CaM-dependency is unknown. In vitro studies have established a complex relationship between calpain cleavage of spectrin, Ca ++ , CaM binding, and the ability of spectrin to oligomerize, bind actin, and bind to membranes (52 , 54).…”

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confidence: 99%

Sequential Degradation of αII and βII Spectrin by Calpain in Glutamate or Maitotoxin-Stimulated Cells

et al. 2006

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Calpain-catalyzed proteolysis of αII-spectrin is a regulated event associated with neuronal long-term potentiation, platelet and leukocyte activation, and other processes. Calpain proteolysis is also linked to apoptotic and non-apoptotic cell death following excessive glutamate exposure, hypoxia, HIVgp120/160 exposure, or toxic injury. The molecular basis for these divergent consequences of calpain action, and their relationship to spectrin proteolysis, is unclear. Calpain preferentially cleaves αII spectrin in vitro in repeat 11 between residues Y 1176 and G 1177 . Unless stimulated by Ca ++ and calmodulin (CaM), βII spectrin proteolysis in vitro is much slower. We identify additional unrecognized sites in spectrin targeted by calpain in vitro and in vivo. Bound CaM induces a second αII spectrin cleavage at G 1230 *S 1231 . βII spectrin is cleaved at four sites. One cleavage only occurs in the absence of CaM at high enzyme-to-substrate ratios near the βII spectrin COOH-terminus. CaM promotes βII spectrin cleavages at Q1440*S1441, S1447*Q1448, and L1482*A1483. These sites are also cleaved in the absence of CaM in recombinant βII spectrin fusion peptides, indicating that they are probably shielded in the spectrin heterotetramer and become exposed only after CaM binds αII spectrin. Using epitope-specific antibodies prepared to the calpain cleavage sites in both αII and βII spectrin, we find in cultured rat cortical neurons that brief glutamate exposure (a physiologic ligand) rapidly stimulates αII spectrin cleavage only at Y 1176 *G 1177 , while βII spectrin remains intact. In cultured SH-SY5Y cells that lack an NMDA receptor, glutamate is without effect. Conversely, when stimulated by calcium influx (via maitotoxin), there is rapid and sequential cleavage of αII and then βII spectrin, coinciding with the onset of non-apoptotic cell death. These results identify: i) novel calpain target sites in both αII and βII spectrin; ii) trans-regulation of proteolytic susceptibility between the spectrin subunits in vivo; and iii) the preferential cleavage of αII spectrin vs. βII spectrin when responsive cells are stimulated by engagement of the NMDA receptor. We postulate that calpain proteolysis of spectrin can activate two physiologically distinct responses: one that enhances skeletal plasticity without destroying the spectrin-actin skeleton, characterized by preservation of βII spectrin; or an alternative response closely correlated with nonapoptotic cell death and characterized by proteolysis of βII spectrin and complete dissolution of the spectrin skeleton. Spectrin is the major component of the cytoskeletal network associated with the plasma membrane of vertebrate cells (for reviews 1 , 2 , 3). While seven spectrin genes exist, encoding two variants of an alpha spectrin and five beta spectrins, the most common form of this protein is a heterotetramer of αII,βII spectrin. Together with actin and a host of adapter proteins, spectrin controls the distribution of many integral and peripheral membrane proteins, and ...

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“…There was also an indication that the interaction of spectrin with the PS-containing monolayer was Ca 2+ dependent [66,81], but this possibility was never explored further. It is possible that complexation of Ca 2+ would affect PS-binding as is known for annexin-type proteins [82,83], or have an effect on spectrin conformation and its sensitivity to calpains [25,84,85].…”

Section: Spectrin Binds the Phospholipid Bilayermentioning

confidence: 99%

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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Calpain‐induced proteolysis of β‐spectrins

Cited by 32 publications

References 31 publications

Regulation of Phosphatidylserine Exposure in Red Blood Cells

Regulation of Phosphatidylserine Exposure in Red Blood Cells

Sequential Degradation of αII and βII Spectrin by Calpain in Glutamate or Maitotoxin-Stimulated Cells

Spectrin and phospholipids — the current picture of their fascinating interplay

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