Human erythrocyte protein 4.1 is a phosphatidylserine binding protein.

Rybicki, Anne C.; Heath, RH; Lubin, Bertram H.; Schwartz, RS

doi:10.1172/jci113303

Cited by 79 publications

(32 citation statements)

References 40 publications

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“…This is in contrast to earlier findings of Cohen et al (24), who obtained a K D value of 3.3 ϫ 10 Ϫ7 M for 4.1R binding to PS. It should also be noted that our binding data are consistent with the data of Rybicki et al (23).…”

Section: Discussionsupporting

confidence: 93%

“…Protein 4.1R has also been shown to bind phosphatidylserine (PS) (22)(23)(24), which is exclusively localized in the inner leaflet of the erythrocyte membrane. However, in contrast to extensive structural and functional characterization of the various protein-protein interactions involving 4.1R, little is known about the nature and the function of 4.1R interaction with PS.…”

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confidence: 99%

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Structural and Functional Characterization of Protein 4.1R-Phosphatidylserine Interaction

An¹,

Takakuwa²,

Manno³

et al. 2001

Journal of Biological Chemistry

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Erythrocyte protein 4.1R is a multifunctional protein that binds to various membrane proteins and to phosphatidylserine. In the present study, we report two important observations concerning 4.1R-phosphatidylserine interaction. Biochemically, a major finding of the present study is that 4.1R binding to phosphatidylserine appears to be a two-step process in which 4.1R first interacts with serine head group of phosphatidylserine through the positively charged amino acids YKRS and subsequently forms a tight hydrophobic interaction with fatty acid moieties. 4.1R failed to dissociate from phosphatidylserine liposomes under high ionic strength but could be released specifically by phospholipase A 2 but not by phospholipase C or D. Biochemical analyses showed that acyl chains were associated with 4.1R released by phospholipase A 2 . Importantly, the association of acyl chains with 4.1R impaired its ability to interact with calmodulin, band 3, and glycophorin C. Removal of acyl chains restored 4.1R binding. These data indicate that acyl chains of phosphatidylserine play an important role in its interaction with 4.1R and on 4.1R function. In terms of biological significance, we have obtained evidence that 4.

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Section: Discussionsupporting

confidence: 93%

mentioning

confidence: 99%

Structural and Functional Characterization of Protein 4.1R-Phosphatidylserine Interaction

An¹,

Takakuwa²,

Manno³

et al. 2001

Journal of Biological Chemistry

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“…Other proteins such as adducin, tropomyosin, tropomodulin, and dematin function as accessory proteins of spectrin-actin junctions and are probably involved in the stabilization of spectrin-actin complexes (36). Furthermore, spectrin and protein 4.1 interact through phosphatidylserine with the inner leaflet of the lipid bilayer (37).…”

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confidence: 99%

Identification of ä-Spectrin Domains Susceptible to Ubiquitination

Corsi

Galluzzi

Lecomte

et al. 1997

Journal of Biological Chemistry

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Previously, we demonstrated that ␣-spectrin is a substrate for the ubiquitin system and that this conjugation is a dynamic process (Corsi, D., Galluzzi, L., Crinelli, R., and Magnani, M. (1995) J. Biol. Chem. 270, 8928 -8935). In this study, we mapped the sites of ubiquitination on erythrocyte ␣-spectrin. A peptide map of digested ␣-spectrin, previously submitted to in vitro 125 I-ubiquitin conjugation, revealed the presence of four distinct labeled bands with M r 40,000, 36,000, 29,000, and 25,500. Western blotting experiments using antibodies against each ␣-spectrin domain revealed that only IgG anti-␣III domain recognized the 125 I-labeled ubiquitin peptide of 29 kDa, whereas the IgG anti-␣V domain recognized the M r 40,000 125 I-ubiquitin-labeled peptide. The other two labeled bands of M r 36,000 and M r 25,500 were identified as tetra and tri multiubiquitin chains. Ubiquitination of the ␣III and ␣V domains was further confirmed by anti-␣-spectrin domain immunoaffinity chromatography. Endoprotease Lys C-digested spectrin conjugated previously to 125 I-ubiquitin was incubated with antibodies against each trypsin-resistant domain of ␣-spectrin. Gamma counting of the radiolabeled antigen-antibody complexes purified by protein A chromatography showed labeling in the IgG anti-␣III and anti-␣V complexes alone. Domain ␣III is not associated with any known function, whereas domain ␣V contains the nucleation site for the association of the ␣ and ␤ chains. Ubiquitination of the latter domain suggests a role for ubiquitin in the modulation of the stability, deformability, and viscoelastic properties of the erythrocyte membrane.

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“…1). This may reflect self-association of protein 4.1 [22] on the cytoskeleton, the attachment of a second protein 4.1 molecule to spectrin to form a 2: 1 complex [23], or binding of protein 4.1 to band 3 [24] or phosphatidylserine [25]. Integrated spectrin staining To determine whether binding of spectrin to a system containing partially saturated actin protofilaments can be observed and reaches a plateau at the total spectrin concentration characteristic of the in vivo complex, we attempted to bring about partial depletion of spectrin by extraction of ghosts at low ionic strength.…”

Section: Resultsmentioning

confidence: 99%

Composition of the ternary protein complex of the red cell membrane cytoskeleton

Pekrun

Pinder

Morris

et al. 1989

European Journal of Biochemistry

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The red cell membrane skeletal network is constructed from actin, spectrin and protein 4.1 in a molar ratio of actin subunits/spectrin heterodimer/protein 4.1 of 2:1:1. This represents saturation of the actin filaments, since incubation with extraneous spectrin and protein 4.1 leads to do binding of additional spectrin, either to the inner surface of ghost membranes or to lipid‐free membrane cytoskeletons. Partial extraction of spectrin from the membrane is accompanied by release of actin under all conditions. Regardless of the proportion of spectrin extracted, the molar ratio of spectrin dimer/actin subunits is constant at 1:2. This is not the result of release or cooperative breakdown of whole lattice junctions from the network, for the number of actin filaments, judged by capacity to nucleate polymerisation of added G‐actin, remains unchanged even when as much as 60% of the total spectrin has been lost. A similar 1:2:1 stoichiometry characterises the complex formed when G‐actin is allowed to polymerise in the presence of varying amounts of spectrin and protein 4.1. When this complex is treated with the depolymerising agent, 1 M guanidine hydrochloride, it breaks down into smaller units of the same stoichiometry. After cross‐linking these can be recovered from a gel‐filtration column. Complexes prepared starting from G‐actin appear to be much more stable than those formed when spectrin and protein 4.1 are bound to F‐actin.

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Human erythrocyte protein 4.1 is a phosphatidylserine binding protein.

Cited by 79 publications

References 40 publications

Structural and Functional Characterization of Protein 4.1R-Phosphatidylserine Interaction

Structural and Functional Characterization of Protein 4.1R-Phosphatidylserine Interaction

Identification of ä-Spectrin Domains Susceptible to Ubiquitination

Composition of the ternary protein complex of the red cell membrane cytoskeleton

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