Electron microscopic evidence of impaired intramembrane particles and instability of the cytoskeletal network in band 4.2 deficiency in human red cells

Yawata, Yoshihito; Yawata, Ayumi; Kanzaki, Akio; Inoue, Takafumi; Okamoto, Naoto; Uehira, Kenzo; Yasunaga, Mutsumi; Nakamura, Yoshihisa

doi:10.1002/(sici)1097-0169(1996)33:2<95::aid-cm3>3.0.co;2-h

Cited by 15 publications

(27 citation statements)

References 25 publications

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“…QFDE was applied for transmission EM as described previously (42). Red cell ghosts were frozen rapidly to Ϫ196ЊC with a quick-freeze apparatus (MF-2; Eiko Co., Tokyo, Japan).…”

Section: Electron Microscopic Analysesmentioning

confidence: 99%

Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation.

Inaba¹,

Yawata²,

Koshino³

et al. 1996

J. Clin. Invest.

169

104

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We studied bovine subjects that exhibited a moderate uncompensated anemia with hereditary spherocytosis inherited in an autosomal incompletely dominant mode and retarded growth. Based on the results of SDS-PAGE, immunoblotting, and electron microscopic analysis by the freeze fracture method, we show here that the proband red cells lacked the band 3 protein completely. Sequence analysis of the proband band 3 cDNA and genomic DNA showed a C → T substitution resulting in a nonsense mutation (CGA → TGA; Arg → Stop) at the position corresponding to codon 646 in human red cell band 3 cDNA. The proband red cells were deficient in spectrin, ankyrin, actin, and protein 4.2, resulting in a distorted and disrupted membrane skeletal network with decreased density. Therefore, the proband red cell membranes were extremely unstable and showed the loss of surface area in several distinct ways such as invagination, vesiculation, and extrusion of microvesicles, leading to the formation of spherocytes. Total deficiency of band 3 also resulted in defective Cl Ϫ /HCO 3 Ϫ exchange, causing mild acidosis with decreases in the HCO 3

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“…QFDE was applied for transmission EM as described previously (42). Red cell ghosts were frozen rapidly to Ϫ196ЊC with a quick-freeze apparatus (MF-2; Eiko Co., Tokyo, Japan).…”

Section: Electron Microscopic Analysesmentioning

confidence: 99%

Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation.

Inaba¹,

Yawata²,

Koshino³

et al. 1996

J. Clin. Invest.

169

104

View full text Add to dashboard Cite

show abstract

“…In our study with fluorescence recovery after use of the photobleaching method (FRAP), the immobile fraction of band 3, which constitutes about 60% of the total in normal red cells, was totally absent in complete protein 4.2 deficiency [5,33]. It is also interesting that, in complete deficiency of protein 4.2, the number of intramembrane particles (IMP) has been found to be reduced with a shift to larger sizes, indicating the possibility of increased oligomerization of band 3 molecules in these red cells [67].…”

Section: Binding Properties Of Protein 42 With Other Membrane Proteinsmentioning

confidence: 65%

“…Golan et al [65] reported that protein 4.2-deficient red cells manifested a significant shift from slowly rotating and rotationally immobile populations of band 3 to a rapidly rotating population, consistent with the interpretation that low-affinity binding sites for band 3 are decreased on the membrane skeleton of protein 4.2-deficient red cells, and that free band 3 dimers are preferentially lost in these red cells, leading to a larger average size of intramembrane particles visualized by freeze fracture electron microscopy [5,33,67]. These results suggest that protein 4.2 deficiency primarily acts to destabilize a fraction of band 3 molecules, resulting in loss of band 3 and possibly of the membrane.…”

Section: Rotational Mobility Of Bandmentioning

confidence: 67%

“…On the other hand, in the red cells of protein 4.2 deficiency of the three types, the number of IMPs had decreased to 4,464 ± 353 µm −2 in the Nippon type, 4,625 ± 381 in protein 4.2 Shiga, and 2,975 ± 310 in protein 4.2 Komatsu [67]. The decreased number of IMPs appeared to be derived from a decreased number of IMPs of small size in association with an increased number of IMPs of medium (9−20 nm) and large size (Ͼ21 nm) [67], indicating increased oligomerization of band 3 in situ ( Figure 4A). The significant contribution of protein 4.2 to the biophysical properties of band 3 was proved by utilizing IOVs of normal controls and those of protein 4.2 deficiency [67].…”

Section: Ultrastructure Of Red Cell Membrane In Situ 7151 Abnormalmentioning

confidence: 93%

“…When intact red cells were subjected to electron microscopic studies using the freeze fracture method, the normal number of IMPs was 5,210 ± 389 µm 2 , approximately of which 80% were basically small in size (4−8 nm) [67]. On the other hand, in the red cells of protein 4.2 deficiency of the three types, the number of IMPs had decreased to 4,464 ± 353 µm −2 in the Nippon type, 4,625 ± 381 in protein 4.2 Shiga, and 2,975 ± 310 in protein 4.2 Komatsu [67].…”

Section: Ultrastructure Of Red Cell Membrane In Situ 7151 Abnormalmentioning

confidence: 99%

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Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

2000

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Protein 4.2 (P4.2) is one of the major red cell membrane proteins, which binds to the membrane skeletal network and to the cytoplasmic domain of anion exchanger band 3, and also interacts with ankyrin. P4.2 plays an important role in maintaining the stability and flexibility of red cells by these biophysical functions. Patients with P4.2 deficiency in their red cell membranes suffer from congenital hemolytic anemia with microspherocytosis or the like. Therefore, new information on the structure and function of the P4.2 gene, characteristics of protein 4.2, and the localization of this protein in the red cell membrane ultrastructure are reviewed. Expression of the gene and protein 4.2 in erythroid and nonerythroid tissues is discussed. In addition, the genotype and phenotype of protein 4.2 deficiency are described in humans and in the targeted P4.2 knock-out (4.2 −/− ) mice.

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The state of DNA methylation in the promoter regions of the human red cell membrane protein (band 3, protein 4.2, andβ-spectrin) genes

et al. 2001

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Indrikis Muiznieks 1,5The state of methylation of the 5Ј-CpG-3Ј sites is known to be linked to the regulation of Birgit Schmitz 1 promoter function by modulating DNA-protein interactions and to the structure of chromaGudrun Schell 1 tin. As part of a project to determine methylation patterns in the human genome, the Yoshihito Yawata 2 methylation profiles were examined in genes for the human erythroid membrane proteins; Walter Doerfler 1 protein 4.2 (P4.2), gene (ELB42), band 3 (B3), gene (EPB3), and β-spectrin (β-Sp), gene 1 Institute of Genetics, University (SPTB). The bisulfite protocol of the genomic sequencing method was applied.(1) In the of Cologne, Köln, Germany DNA from peripheral white blood cells, the promoter regions of EPB3 and ELB42 were 2 Division of Hematology, extensively methylated, but the promoter of SPTB was totally unmethylated. (2) During Department of Medicine, erythroid differentiation, (i) ELB42 was unmethylated in DNAs from the cell line UT-7/EPO, Kawasaki Medical School, but became methylated (55Ϫ93%) in cultured erythroblasts from peripheral BFU-E. The Kurashiki City, Japan mRNA from ELB42 was first detected in early erythroblasts and protein 4.2 was expressed Present addresses: in late erythroblasts. (ii) EPB3 was consistently methylated in UT-7/EPO and also in cultu-3 Institute of Human Genetics red erythroblasts from burst forming unit erythroid (BFU-E) from peripheral blood. EPB3 and Anthropology, Heinrichand ELB42 were efficiently transcribed in UT-7 cells only after erythropoietin stimulation. Heine-University, Duesseldorf, (iii) SPTB remained unmethylated in DNAs from UT-7/EPO and cultured erythroblasts. (3) Germany 4 Institute of Human Genetics, We also investigated methylation profiles in peripheral white blood cells from patients with University of Essen Medical erythroid diseases, like complete P4.2 deficiency due to ELB42 mutations, hereditary School, Essen, Germany spherocytosis with EPB3 mutations, and hereditary elliptocytosis with SPTB mutations. 5 Department of Microbiology,The methylation profiles of the promoter regions of these three genes were essentially Latvian University, Riga, Latvia identical to those in healthy individuals.

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Electron microscopic evidence of impaired intramembrane particles and instability of the cytoskeletal network in band 4.2 deficiency in human red cells

Cited by 15 publications

References 25 publications

Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation.

Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation.

Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

The state of DNA methylation in the promoter regions of the human red cell membrane protein (band 3, protein 4.2, andβ-spectrin) genes

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