Identification of the Protein 4.1 Binding Interface on Glycophorin C and p55, a Homologue of the Drosophila discs-large Tumor Suppressor Protein

Marfatia, Shirin M.; Lue, Robert A.; Branton, Daniel; Chishti, Athar H.

doi:10.1074/jbc.270.2.715

Cited by 200 publications

(199 citation statements)

References 36 publications

Supporting

Mentioning

191

Contrasting

Unclassified

Order By: Relevance

“…The nucleotide primers required for the cloning of the construct have been described previously (14). The GST-GPC 82-128 protein was affinity-purified on the glutathione-Sepharose beads, and the GPC 82-128 protein was cleaved from GST on the column by thrombin (18).…”

Section: Methodsmentioning

confidence: 99%

“…We have previously shown that p55 directly binds to protein 4.1 and glycophorin C and may form a ternary complex with these proteins at the cytoplasmic face of the plasma membrane (14). This ternary complex links the spectrin-actin junctions to the lipid bilayer and is presumed to play an important role in maintaining erythrocyte shape and its membrane material properties (15,16).…”

mentioning

confidence: 99%

“…This ternary complex links the spectrin-actin junctions to the lipid bilayer and is presumed to play an important role in maintaining erythrocyte shape and its membrane material properties (15,16). To delineate the basis of p55 interactions with protein 4.1 and glycophorin C, we have mapped the binding interfaces between protein 4.1 and p55 as well as between protein 4.1 and glycophorin C (17). However, the site where the cytoplasmic domain of glycophorin C binds to p55 had not yet been determined.…”

mentioning

confidence: 99%

See 2 more Smart Citations

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

Self Cite

View full text Add to dashboard Cite

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...

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…The founding member of the protein 4.1 family, 4.1R (official mouse protein symbol, Epb4.1), was originally identified as a stabilizer of erythrocyte shape, connecting the transmembrane protein glycophorin C (GLPC) and the MAGUK p55 with the spectrin cytoskeleton (Marfatia et al, 1995). In addition to 4.1R, AMPA-type glutamate receptors mediate fast excitatory synaptic transmission in the vertebrate brain.…”

Section: Introductionmentioning

confidence: 99%

The function of glutamatergic synapses is not perturbed by severe knockdown of 4.1N and 4.1G expression

Wozny

Breustedt

Wolk

et al. 2009

Journal of Cell Science

View full text Add to dashboard Cite

AMPA-type glutamate receptors mediate fast excitatory synaptic transmission in the vertebrate brain. Their surface expression at synapses between neurons is regulated in an activity-dependent and activity-independent manner. The protein machinery that regulates synaptic targeting, anchoring and turnover of AMPA receptors consists of several types of specialized scaffolding proteins. The FERM domain scaffolding proteins 4.1G and 4.1N were previously suggested to act jointly in binding and regulating synaptic trafficking of the AMPA receptor subunits GluR1 and GluR4. To determine the functions of 4.1G and 4.1N in vivo, we generated a mutant mouse line that lacks 4.1G entirely and expresses 4.1N at 22% of wild-type levels. These mice had combined 4.1G and 4.1N protein expression in the hippocampus at 12% of wild-type levels (equivalent to 8-10% of combined GluR1 and GluR4 expression levels). They show a moderate reduction in synaptosomal expression levels of the AMPA receptor subunit GluR1 at 3 weeks of age, but no change in basic glutamatergic synaptic transmission and long-term potentiation in the hippocampus. Our study indicates that 4.1G and 4.1N do not have a crucial role in glutamatergic synaptic transmission and the induction and maintenance of long-term plastic changes in synaptic efficacy.

show abstract

“…Based on amino acid sequencing, c-merlin possesses several protein motifs highly similar to merlin molecules identified in other species, including a 300-residue FERM domain located at the N-terminal end of chicken merlin. FERM domains in other Protein 4.1 molecules have been shown to mediate interactions with cell surface glycoproteins, including CD44, glycophorin-C, and p55 (Marfatia et al, 1995;Hirao et al, 1996). The interaction between merlin and CD44 may have functional importance in transducing growth arrest signals (Morrison et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Characterization of chicken Nf2/merlin indicates regulatory roles in cell proliferation and migration

Chen

Gutmann

Haipek

et al. 2004

Developmental Dynamics

View full text Add to dashboard Cite

The neurofibromatosis 2 (NF2) tumor suppressor protein merlin, or schwannomin, functions as a negative growth regulator such that inactivating mutations in Nf2 predispose humans to tumors. In addition, merlin has a critical role during embryonic development. Nf2-deficient mice die early during embryogenesis, with defects in gastrulation and extraembryonic tissues. To investigate the function of Nf2/merlin during embryonic development, we first identified the homologous Nf2 gene in chicken (cNf2) and examined the distribution of chicken merlin (c-merlin) during myogenesis. cNf2 encoded a full-length mRNA of 1,770 nucleotides and a protein of 589 residues. C-merlin shared high sequence homology and common protein motifs with vertebrate and Drosophila merlins. In addition, cNF2 functions as a negative growth regulator similar to human and Drosophila merlin in vitro. In vivo, c-merlin was expressed diffusely in the forming dermomyotome but down-regulated in migratory muscle precursors in the forelimb. As muscle formed in the limb, c-merlin expression was up-regulated. As an initial examination of c-merlin function during myogenesis, c-merlin was ectopically expressed in muscle precursors and the effects on muscle development were examined. We show that ectopic merlin expression reduces the proliferation of muscle precursors as well as their ability to migrate effectively in limb mesoderm. Collectively, these results demonstrate that c-merlin is developmentally regulated in migrating and differentiating myogenic cells, where it functions as a negative regulator of both muscle growth and motility. Developmental Dynamics 229:541-554, 2004.

show abstract

Identification of the Protein 4.1 Binding Interface on Glycophorin C and p55, a Homologue of the Drosophila discs-large Tumor Suppressor Protein

Cited by 200 publications

References 36 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

The function of glutamatergic synapses is not perturbed by severe knockdown of 4.1N and 4.1G expression

Characterization of chicken Nf2/merlin indicates regulatory roles in cell proliferation and migration

Contact Info

Product

Resources

About