Restoration of normal membrane stability to unstable protein 4.1-deficient erythrocyte membranes by incorporation of purified protein 4.1.

Takakuwa, Yuichi; Tchernia, Gil; Rossi, Mary; Benabadji, M.; Mohandas, Narla

doi:10.1172/jci112577

Cited by 105 publications

(74 citation statements)

References 32 publications

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“…This hypothesis assumes that only a subpopulation of protein 4.1 regulates SSPC at the plasma membrane. The remaining protein 4.1 may independently regulate properties such as membrane deformability, which is restored by the addition of exogenous protein 4.1 in the absence of either p55 or hDlg (43,44).…”

Section: Discussionmentioning

confidence: 99%

The PDZ Domain of Human Erythrocyte p55 Mediates Its Binding to the Cytoplasmic Carboxyl Terminus of Glycophorin C

Marfatia

Morais-Cabral²,

Kim

et al. 1997

Journal of Biological Chemistry

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The PDZ domain, also known as the GLGF repeat/DHR domain, is an ϳ90-amino acid motif discovered in a recently identified family of proteins termed MAGUKs (membrane-associated guanylate kinase homologues). Sequence comparison analysis has since identified PDZ domains in over 50 proteins. Like SH2 and SH3 domains, the PDZ domains mediate specific protein-protein interactions, whose specificities appear to be dictated by the primary structure of the PDZ domain as well as its binding target. Using recombinant fusion proteins and a blot overlay assay, we show that a single copy of the PDZ domain in human erythrocyte p55 binds to the carboxyl terminus of the cytoplasmic domain of human erythroid glycophorin C. Deletion mutagenesis of 21 amino acids at the amino terminus of the p55 PDZ domain completely abrogates its binding activity for glycophorin C. Using an alanine scan and surface plasmon resonance technique, we identify residues in the cytoplasmic domain of glycophorin C that are critical for its interaction with the PDZ domain. The recognition specificity of the p55 PDZ domain appears to be unique, since the three PDZ domains of hDlg (human lymphocyte homologue of the Drosophila discs large tumor suppressor) do not bind the cytoplasmic domain of glycophorin C. Taken together with our previous studies, these results complete the identification of interacting domains in the ternary complex between p55, glycophorin C, and protein 4.1. Implications of these findings are discussed in terms of binding specificity and the regulation of cytoskeleton-membrane interactions.p55 is a heavily palmitoylated peripheral membrane protein of the red blood cell membrane (1). The primary structure of p55 defines several distinct domains including the PDZ domain, the SH3 domain, the protein 4.1 binding domain, the tyrosine phosphorylation domain, and a carboxyl-terminal guanylate kinase-like domain (2). The domain organization of p55 is similar to a family of proteins termed MAGUKs 1 (membraneassociated guanylate kinase homologues), which appear to play important roles in the regulation of signaling pathways and cytoskeleton-membrane interactions (3-6). Among various protein domains of MAGUKs, the PDZ domain has been a focus of intense scrutiny (5-8). Like the SH2 and SH3 domains, the PDZ domains recruit cytoplasmic proteins to specific submembranous sites. For example, the PDZ 1 ϩ 2 domain, but not the PDZ 3 domain, of PSD-95 and hDlg interacts with the carboxyl terminus of the NMDA receptors and the Shaker type K ϩ channels, and this interaction plays an important role in the clustering of ion channels (5, 6). The PDZ domains have also been shown to interact with other PDZ domains. The PDZ domain of neuronal nitric-oxide synthase binds to the PDZ domain of syntrophin at the sarcolemma in skeletal muscle and to the PDZ 2 domain of PSD-95 at the synaptic junctions of neuronal cells (9, 10). These observations suggest that the primary structures of both the PDZ domain and its target peptide govern the specificity as well as the affin...

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Section: Discussionmentioning

confidence: 99%

The PDZ Domain of Human Erythrocyte p55 Mediates Its Binding to the Cytoplasmic Carboxyl Terminus of Glycophorin C

Marfatia

Morais-Cabral²,

Kim

et al. 1997

Journal of Biological Chemistry

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“…By interacting with components of the spectrin-based membrane skeleton, as well as with integral proteins in the overlying lipid bilayer, 4.1R is critical to both the structural integrity of the skeleton and its attachment to the membrane. Deficiency of 4.1R results in red cells with morphological abnormalities and unstable membranes due to the assembly of a mechanically compromised membrane skeleton (1,2). Such defects are manifested by the appearance of circulating cells with elliptical morphology and also by the presence of fragmented cells due to the decreased membrane mechanical stability.…”

Section: Introductionmentioning

confidence: 99%

Protein 4.1R–deficient mice are viable but have erythroid membrane skeleton abnormalities

et al. 1999

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A diverse family of protein 4.1R isoforms is encoded by a complex gene on human chromosome 1. Although the prototypical 80-kDa 4.1R in mature erythrocytes is a key component of the erythroid membrane skeleton that regulates erythrocyte morphology and mechanical stability, little is known about 4.1R function in nucleated cells. Using gene knockout technology, we have generated mice with complete deficiency of all 4.1R protein isoforms. These 4.1R-null mice were viable, with moderate hemolytic anemia but no gross abnormalities. Erythrocytes from these mice exhibited abnormal morphology, lowered membrane stability, and reduced expression of other skeletal proteins including spectrin and ankyrin, suggesting that loss of 4.1R compromises membrane skeleton assembly in erythroid progenitors. Platelet morphology and function were essentially normal, indicating that 4.1R deficiency may have less impact on other hematopoietic lineages. Nonerythroid 4.1R expression patterns, viewed using histochemical staining for lacZ reporter activity incorporated into the targeted gene, revealed focal expression in specific neurons in the brain and in select cells of other major organs, challenging the view that 4.1R expression is widespread among nonerythroid cells. The 4.1R knockout mice represent a valuable animal model for exploring 4.1R function in nonerythroid cells and for determining pathophysiological sequelae to 4.1R deficiency.

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“…Shi et al (1999) reported that erythrocytes from 4.1R-knockout mice demonstrate membrane instability and reduced levels of other cytoskeletal proteins, consequently leading to hemolytic anemia. In addition, Stagg (Takakuwa et al, 1986;Mohler et al, 2004;Baines et al, 2009) showed that electrocardiographic analysis revealed diminished heart rate with a prolonged Q-T interval and reduction of the Na + /Ca 2+ exchanger current density in 4.1R-deficient (knockout) mice. However, no changes regarding the ejection fraction or fractional shortening have been observed in 4.1R-deficient mice, as assessed by echocardiography (Stagg et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Correlation between protein 4.1R and the progression of heart failure in vivo

Wei

Yang

et al. 2016

Genet. Mol. Res.

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ABSTRACT. We aimed to assess the protein 4.1R (4.1R) expression of the membrane skeleton in cardiomyocytes and to determine the potential role of 4.1R in the pathogenesis of heart failure (HF). Forty-two male mice were randomly divided into two groups: an HF group (N = 22) and control group (N = 20). The HF model was established by abdominal subcutaneous injection of 5 mg⋅kg -1 ⋅day -1 isopropyl adrenaline to the mice for 14 days. Electrocardiography was carried out and cardiac function was assessed by ultrasonic cardiogram. The left ventricular weight index (LVMI) was measured after mice were sacrificed, and the pathological changes of the heart were observed by hematoxylin and eosin staining. The expression of 4.1R in cardiomyocytes was analyzed by immunohistochemistry, immunofluorescence, and reverse transcription polymerase chain reaction. The echocardiographs showed that the left ventricular end-diastolic dimension (LVEDD) and left ventricular end-systolic dimension (LVESD) were significantly higher in the HF group than in the controls (P < 0.05), while the left ventricle shortening fraction was remarkably lower than that in the controls (P < 0.05). Electrocardiography showed faster heart rates in the HF group than in the control group (P < 0.05). Both the LVMI and the myocardial tissue pathological score were significantly higher (P < 0.01) in the HF group than in the controls. 4.1R localized mostly to the plasma membrane and was distributed discretely in the cytosol of myocardial cells. The proportion of 4.1R-positive cells was significantly higher in the HF group (P < 0.01) than in the controls, which was confirmed by the positive mRNA expression of 4.1R. 4.1R localized mostly to the plasma membrane of myocardial cells and was upregulated with the progression of HF. This suggests that 4.1R may be associated with HF progression and therefore 4.1R represents a promising therapeutic target in HF.

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Restoration of normal membrane stability to unstable protein 4.1-deficient erythrocyte membranes by incorporation of purified protein 4.1.

Cited by 105 publications

References 32 publications

The PDZ Domain of Human Erythrocyte p55 Mediates Its Binding to the Cytoplasmic Carboxyl Terminus of Glycophorin C

The PDZ Domain of Human Erythrocyte p55 Mediates Its Binding to the Cytoplasmic Carboxyl Terminus of Glycophorin C

Protein 4.1R–deficient mice are viable but have erythroid membrane skeleton abnormalities

Correlation between protein 4.1R and the progression of heart failure in vivo

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