Evidence that the red cell skeleton protein 4.2 interacts with the Rh membrane complex member CD47

Mouro-Chanteloup, Isabelle; Delaunay, Jean; Gane, Pierre; Nicolas, Virginie; Johansen, Mette L.; Brown, Eric J.; Peters, Luanne L.; Kim, Caroline Le Van; Cartron, Jean Pierre; Colin, Yves

doi:10.1182/blood-2002-04-1285

Cited by 118 publications

(142 citation statements)

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“…Because Rh and RhAG deficiency is associated with stomato-spherocytosis of Rh null red cells, we and others have analyzed Rh complex expression in some HS red cells with primary defects in different skeletal or transmembrane proteins. Using this approach, a positive correlation between the expression levels of CD47 and protein 4.2 has been recently reported from the analysis of 4.2(Ϫ)HS red cells (21,22). However, as discussed in our introduction, the lack of protein 4.2 has no deleterious effect on the expression level and skeleton linkage of Rh and RhAG.…”

Section: The Rh Null -Associated Substitution Of Asp-399 Alters Intermentioning

confidence: 99%

“…Preparation of RBC membranes, SDS-PAGE, and Western blotting of membrane proteins were done as described previously (22). For flow cytometric measurement, 2 l of packed red cells were fixed at room temperature in 1 ml of 1% formol/0.025% glutaraldehyde for 15 min and then in 1 ml of 3% formol overnight.…”

Section: Methodsmentioning

confidence: 99%

“…Are Rh-and RhAG-deficient-Cell surface expression of the Rh complex on RBCs from ankyrin-R-deficient (nb/nb) and ␤-adducin-deficient (adducin Ϫ/Ϫ ) mice was compared with wild type by flow cytometric analysis of Rh, RhAG, and CD47, as described previously (22). This analysis revealed that Rh and RhAG were reduced by 53 and 97%, respectively, in RBCs from nb/nb mice (n ϭ 3) compared with wild type and adducin Ϫ/Ϫ , whereas CD47 was normally expressed in all samples (Fig.…”

Section: Ankyrin-r-deficient Mouse Rbcsmentioning

confidence: 99%

“…Recently, an association between CD47 and protein 4.2 was indicated by the observation that RBCs with complete protein 4.2 deficiency (4.2(Ϫ)HS) exhibit a 80 -90% reduction of CD47 (21,22). However, CD47 linkage to the red cell skeleton was found to be independent of Rh and RhAG (20), and neither the expression level nor the Triton X-100 extraction of Rh and RhAG from the cell membrane were altered by the absence of protein 4.2 in 4.2(Ϫ)HS RBCs (22). These results strongly suggest that protein 4.2 is not involved in linking the core of the Rh complex to the membrane skeleton.…”

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confidence: 99%

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Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation

Nicolas

Kim

Gane

et al. 2003

Journal of Biological Chemistry

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Several studies suggest that the Rh complex represents a major interaction site between the membrane lipid bilayer and the red cell skeleton, but little is known about the molecular basis of this interaction. We report here that ankyrin-R is capable of interacting directly with the C-terminal cytoplasmic domain of Rh and RhAG polypeptides. We first show that the primary defect of ankyrin-R in normoblastosis (nb/nb) spherocytosis mutant mice is associated with a sharp reduction of RhAG and Rh polypeptides. Secondly, our flow cytometric analysis of the Triton X-100 extractability of recombinant fusion proteins expressed in erythroleukemic cell lines suggests that the C-terminal cytoplasmic domains of Rh and RhAG are sufficient to mediate interaction with the erythroid membrane skeleton. Using the yeast two-hybrid system, we demonstrate a direct interaction between the cytoplasmic tails of Rh and RhAG and the second repeat domain (D2) of ankyrin-R. This finding is supported by the demonstration that the substitution of Asp-399 in the cytoplasmic tail of RhAG, a mutation associated with the deficiency of the Rh complex in one Rh null patient, totally impaired interaction with domain D2 of ankyrin-R. These results identify the Rh/RhAG-ankyrin complex as a new interaction site between the red cell membrane and the spectrin-based skeleton, the disruption of which might result in the stomato-spherocytosis typical of Rh null red cells.

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Section: The Rh Null -Associated Substitution Of Asp-399 Alters Intermentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Ankyrin-r-deficient Mouse Rbcsmentioning

confidence: 99%

mentioning

confidence: 99%

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Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation

Nicolas

Kim

Gane

et al. 2003

Journal of Biological Chemistry

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118

109

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“…Although CD47 -SIRP ␣ interactions appear to de-activate autologous macrophages in mouse, severe reductions of CD47 (perhaps 90%) are found on human blood cells from some Rh genotypes who show little to no evidence of anemia ( Mouro-Chanteloup et al, 2003 ) and also little to no evidence of enhanced cell interactions with phagocytic monocytes ( Arndt and Garratty, 2004 ). Here, we assess the species-specifi c inhibition of phagocytosis by CD47 -SIRP ␣ interactions at the phagocytic synapse and visualize how the interaction affects cytoskeletal activity.…”

Section: Introductionmentioning

confidence: 99%

Inhibition of “self” engulfment through deactivation of myosin-II at the phagocytic synapse between human cells

Tsai¹,

Discher²

2008

J Exp Med

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(2008). Inhibition of "self " engulfment through deactivation of myosin-II at the phagocytic synapse between human cells. Retrieved from http://repository.upenn.edu/cbe_papers/108Inhibition of "self " engulfment through deactivation of myosin-II at the phagocytic synapse between human cells AbstractPhagocytosis of foreign cells or particles by macrophages is a rapid process that is inefficient when faced with "self " cells that display CD47-although signaling mechanisms in self-recognition have remained largely unknown. With human macrophages, we show the phagocytic synapse at cell contacts involves a basal level of actin-driven phagocytosis that, in the absence of species-specific CD47 signaling, is made more efficient by phospho-activated myosin. We use "foreign" sheep red blood cells (RBCs) together with CD47-blocked, antibody-opsonized human RBCs in order to visualize synaptic accumulation of phosphotyrosine, paxillin, Factin, and the major motor isoform, nonmuscle myosin-IIA. When CD47 is functional, the macrophage counter-receptor and phosphatase-activator SIRPα localizes to the synapse, suppressing accumulation of phosphotyrosine and myosin without affecting F-actin. On both RBCs and microbeads, human CD47 potently inhibits phagocytosis as does direct inhibition of myosin. CD47-SIRPα interaction initiates a dephosphorylation cascade directed in part at phosphotyrosine in myosin. A point mutation turns off this motor's contribution to phagocytosis, suggesting that self-recognition inhibits contractile engulfment. Comments

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Red Cell Membrane Abnormalities

Gallagher¹

2006

Pediatric Hematology

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Evidence that the red cell skeleton protein 4.2 interacts with the Rh membrane complex member CD47

Cited by 118 publications

References 40 publications

Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation

Rh-RhAG/Ankyrin-R, a New Interaction Site between the Membrane Bilayer and the Red Cell Skeleton, Is Impaired by Rhnull-associated Mutation

Inhibition of “self” engulfment through deactivation of myosin-II at the phagocytic synapse between human cells

Red Cell Membrane Abnormalities

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