The spectrin repeat folds into a three‐helix bundle in solution

Pascual, Jaime; Pfuhl, Mark; Rivas, Germán; Pastore, Annalisa; Saraste, Matti

doi:10.1016/0014-5793(96)00251-7

Cited by 69 publications

(48 citation statements)

References 44 publications

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“…8). A similar behavior of loops has been observed in other proteins, e.g., interleukin-4 (Redfield et al., 1992) or the spectrin repeat (Pascual et al, 1996). The rather compact fold and the presence of the heme as a covalent linker may prevent a higher mobility of the loops.…”

Section: Mobilitysupporting

confidence: 70%

NMR assignments and relaxation studies of Thiobacillus versutus ferrocytochrome c‐550 indicate the presence of a highly mobile 13‐residues long C‐terminal tail

Ubbink¹,

Pfuhl

Oost

et al. 1996

Protein Science

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Cytochrome c-550 of Thiobacillus versutus functions as an electron transfer protein in a chain of redox proteins that enables I: versutus to grow on methylamine. It is a single-heme protein of 134 residues, related to mitochondrial cytochrome c. Cytochrome c-550, as well as several other bacterial c2-type cytochromes, contain a C-terminal extension of 13-16 amino acids of unknown function, compared to mitochondrial cytochrome c.NMR experiments were performed to obtain structural and dynamic information on the protein in solution. For this purpose, I: versutus cytochrome c-550 was labeled with '*N and I3C using I3C-methanol grown Paracoccus denitrificans as a host for heterologous expression. NMR assignments were obtained for the 'H, "N, and I3C nuclei in the backbone and the P-positions of the protein and the secondary structure was determined. "N-relaxation studies were performed to characterize the dynamic properties of the protein.The results indicate that the main part of I: versutus ferrocytochrome c-550 exists in solution as a rigid, well-ordered molecule with a secondary structure that is very similar to that of P. denitrificans cytochrome c-550, as observed in crystals. The C-terminal extension, however, is unstructured and highly mobile. The possible origin and function of the extension are discussed.

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Section: Mobilitysupporting

confidence: 70%

NMR assignments and relaxation studies of Thiobacillus versutus ferrocytochrome c‐550 indicate the presence of a highly mobile 13‐residues long C‐terminal tail

Ubbink¹,

Pfuhl

Oost

et al. 1996

Protein Science

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“…2 Refinements and phasing of the model have been made as determination of the primary structure, 3,4 X-ray, nuclear magnetic resonance, and spectroscopic studies of spectrin have been completed. 28,[35][36][37][38][39][40][41][42][43] These studies predict that the first and third helices are parallel and the second helix is antiparallel. The amphipathic repeats are stabilized by hydrophobic interactions of each repeat at the interior core of each repeat.…”

Section: Discussionmentioning

confidence: 99%

Mutation of a highly conserved isoleucine disrupts hydrophobic interactions in the αβ spectrin self-association binding site

et al. 2003

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We studied an infant with severe neonatal hemolytic anemia and hyperbilirubinemia that evolved into a partially compensated ellipto-poikilocytic anemia. His father had typical elliptocytosis. Their erythrocyte membranes demonstrated structural and functional defects in spectrin. Genetic studies revealed that the proband and his father were heterozygous for an a-spectrin mutation, Ile24Thr, in the ab spectrin self-association binding site. The proband also carried the low expression allele a LELY in trans, influencing the clinical phenotype. The importance of isoleucine in this position of the proposed triple helical model of spectrin repeats is highlighted by its evolutionary conservation in all a spectrins from Drosophila to humans. Molecular modeling demonstrated that replacement of a hydrophobic isoleucine with a hydrophilic threonine disrupts highly conserved hydrophobic interactions in the interior of the spectrin triple helix critical for spectrin function.

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“…The presence of SRs in the plakin domain was confirmed by the crystal structures of two di-repeat fragments of BPAG1 (29) and plectin (30). The SR-fold (ϳ100-residues long) consists of three ␣-helices connected by short loops that pack in a left-handed helical bundle with up-down-up topology (31)(32). The ␣-helices show a heptad pattern in which positions a and d are occupied by hydrophobic residues that pack at the core of the bundle.…”

mentioning

confidence: 88%

The Structure of the Plakin Domain of Plectin Reveals a Non-canonical SH3 Domain Interacting with Its Fourth Spectrin Repeat

Ortega

Buey

Sonnenberg

et al. 2011

Journal of Biological Chemistry

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Plectin belongs to the plakin family of cytoskeletal crosslinkers, which is part of the spectrin superfamily. Plakins contain an N-terminal conserved region, the plakin domain, which is formed by an array of spectrin repeats (SR) and a Src-homology 3 (SH3), and harbors binding sites for junctional proteins. We have combined x-ray crystallography and small angle x-ray scattering (SAXS) to elucidate the structure of the central region of the plakin domain of plectin, which corresponds to the SR3, SR4, SR5, and SH3 domains. The crystal structures of the SR3-SR4 and SR4-SR5-SH3 fragments were determined to 2.2 and 2.95 Å resolution, respectively. The SH3 of plectin presents major alterations as compared with canonical Pro-rich binding SH3 domains, suggesting that plectin does not recognize Pro-rich motifs. In addition, the SH3 binding site is partially occluded by an intramolecular contact with the SR4. Residues of this pseudobinding site and the SR4/SH3 interface are conserved within the plakin family, suggesting that the structure of this part of the plectin molecule is similar to that of other plakins. We have created a model for the SR3-SR4-SR5-SH3 region, which agrees well with SAXS data in solution. The three SRs form a semi-flexible rod that is not altered by the presence of the SH3 domain, and it is similar to those found in spectrins. The flexibility of the plakin domain, in analogy with spectrins, might contribute to the role of plakins in maintaining the stability of tissues subject to mechanical stress.Plakins are a family of high molecular weight proteins that interconnect elements of the cytoskeleton and tether them to membrane-associated structures; hence, they are also known as cytolinkers (1-2). Mammalian plakins include desmoplakin, plectin, the bullous pemphigoid antigen 1 (BPAG1), 4 the microtubule actin crosslinking factor 1 (MACF1, also known as ACF7, trabeculin, or macrophin), envoplakin, periplakin, and epiplakin. Plakins are also present in invertebrates; Drosophila melanogaster has a single plakin gene named shortstop (also know as kakapo) that encodes at least two protein forms, Shot I and Shot II. Similarly, the only plakin gene in Caenorhabditis elegans, vab-10, encodes two variants VAB-10A and VAB-10B.Plectin is a highly versatile plakin that associates with intermediate filaments (3), microtubules (4), and actin fibers (5), and crosslinks these cytoskeletal networks (5-6). Plectin also connects intermediate filaments to membrane-associated complexes. In stratified epithelia such as the skin, plectin is localized at the hemidesmosomes, which are junctional complexes that link the intermediate filaments to the basement membrane, and links the integrin ␣6␤4 and the bullous pemphigoid antigen 2 (BPAG2, also known as type XVII collagen or BP180) to the cytokeratins (7-8). Plectin is also localized at the desmosomes (9), which mediate cell-cell contacts, and connects the nuclear envelope to the intermediate filaments by binding to the outer nuclear membrane protein nesprin-3␣ (10 -11). In st...

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The spectrin repeat folds into a three‐helix bundle in solution

Cited by 69 publications

References 44 publications

NMR assignments and relaxation studies of Thiobacillus versutus ferrocytochrome c‐550 indicate the presence of a highly mobile 13‐residues long C‐terminal tail

NMR assignments and relaxation studies of Thiobacillus versutus ferrocytochrome c‐550 indicate the presence of a highly mobile 13‐residues long C‐terminal tail

Mutation of a highly conserved isoleucine disrupts hydrophobic interactions in the αβ spectrin self-association binding site

The Structure of the Plakin Domain of Plectin Reveals a Non-canonical SH3 Domain Interacting with Its Fourth Spectrin Repeat

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