Binding sites of calmodulin and actin on the brain spectrin, calspectin.

Tsukita, Sachiko; Ishikawa, Harunori; Kurokawa, Manae S.; Morimoto, Kouichi; Sobue, Kenji; Kakiuchi, Shiro

doi:10.1083/jcb.97.2.574

Cited by 52 publications

(23 citation statements)

References 20 publications

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“…Such a positioning is supported by Carlin et al (1983) who found that the major proteolytic breakdown product of fodrin (spectrin), which represents the COOH-terminal part of the chain is able to bind calmodulin. Tsukita et al (1983) have, however, suggested that the calmodulin-binding site is close to the amino terminus of the c~-chain. Subcloning and expression of the cDNA containing the putative calmodulinbinding site will enable us to test our proposal.…”

Section: The Nonhomologous Segments As the "Carriers'of The Distinct mentioning

confidence: 99%

“…In the antiparallel orientation of the subunits in ct-actinin dimer, the actin-binding NH2 terminus comes near the COOH terminus of the adjacent subunit which may then affect the binding (Noegel et al, 1987). In spectrin, actin binding occurs at the ends of the tetramers and involves the COOH terminus of the o~-subunit and NH2 terminus of the /~-subunit (Morrow et al, 1980;Tsukita et al, 1983). Analogously to c~-actinin, it can be surmised that the principal actin binding would occur at the NH2 terminus of the B-chain with the COOH terminus of the or-chain exerting control on the interaction.…”

Section: The Nonhomologous Segments As the "Carriers'of The Distinct mentioning

confidence: 99%

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Primary structure of the brain alpha-spectrin [published erratum appears in J Cell Biol 1989 Mar;108(3):following 1175]

Wasenius

1989

The Journal of Cell Biology

157

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Abstract. We have determined the nucleotide sequence coding for the chicken brain a-spectrin. It is derived both from the cDNA and genomic sequences, comprises the entire coding frame, 5' and 3' untranslated sequences, and terminates in the poly(A)-tail. The deduced amino acid sequence was used to map the domain structure of the protein. The a-chain of brain spectrin contains 22 segments of which 20 correspond to the repeat of the human erythrocyte spectrin (Speicher, D. W., and V. T. Marchesi. 1984. Nature (Lond.). 311:177-180.), typically made of 106 residues. These homologous segments probably account for the flexible, rod-like structure of spectrin. Secondary structure prediction suggests predominantly a-helical structure for the entire chain.Parts of the primary structure are excluded from the repetitive pattern and they reside in the middle part of the sequence and in its COOH terminus. Search for homology in other proteins showed the presence of the following distinct structures in these nonrepetitive regions: (a) the COOH-terminal part of the molecule that shows homology with a-actinin, (b) two typical EF-hand (i.e., Ca:*-binding) structures in this region, (c) a sequence close to the EF-hand that fulfills the criteria for a calmodulin-binding site, and (d) a domain in the middle of the sequence that is homologous to a NH2-terminal segment of several src-tyrosine kinases and to a domain of phospholipase C. These regions are good candidates to carry some established as well as some yet unestablished functions of spectrin. Comparative analysis showed that a-spectrin is well conserved across the species boundaries from Xenopus to man, and that the human erythrocyte a-spectrin is divergent from the other spectrins.

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Section: The Nonhomologous Segments As the "Carriers'of The Distinct mentioning

confidence: 99%

Section: The Nonhomologous Segments As the "Carriers'of The Distinct mentioning

confidence: 99%

Primary structure of the brain alpha-spectrin [published erratum appears in J Cell Biol 1989 Mar;108(3):following 1175]

Wasenius

1989

The Journal of Cell Biology

157

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“…Similar isoforms in mice have also been studied [21]. Spectrin II (SpII) is often referred to as brain spectrin, non-erythroid spectrin [22], calspectin [23] or fodrin [24], and is originally found in neuronal axons, but not dendrites [25]. SpI is confined to neuronal cell bodies and dendrites, and some glial cells, but is not present in axons or presynaptic terminals [26].…”

Section: Introductionmentioning

confidence: 99%

Brain proteins interacting with the tetramerization region of non-erythroid alpha spectrin

Fung

2007

Cellular and Molecular Biology Letters

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Abstract:The N-terminal region of non-erythroid alpha spectrin (SpαII) is responsible for interacting with its binding partner, beta spectrin, to form functional spectrin tetramers. We used a yeast-two-hybrid system, with an N-terminal segment of alpha spectrin representing the functional tetramerization site, as a bait to screen human brain c-DNA library for proteins that interact with the alpha spectrin segment. In addition to several beta spectrin isoforms, we identified 14 proteins that interact with SpαII. Seven of the 14 were matched to 6 known proteins: Duo protein, Lysyl-tRNA synthetase, TBP associated factor 1, two isoforms (b and c) of a protein kinase A interacting protein and Zinc finger protein 333 (2 different segments). Four of the 6 proteins are located primarily in the nucleus, suggesting that spectrin plays important roles in nuclear functions. The remaining 7 proteins were unknown to the protein data base. Structural predictions show that many of the 14 proteins consist of a large portion of unstructured regions, suggesting that many of these proteins fold into a rather flexible conformation. It is interesting to note that all but 3 of the 14 proteins are predicted to consist of one to four coiled coils (amphiphilic helices). A mutation in SpαII, V22D, which interferes with the coiled coil bundling of SpαII with beta spectrin, also affects SpαII interaction with Duo protein, TBP associated factor 1 and Lysyl-tRNA synthetase, suggesting that they may compete with beta spectrin for interaction with SpαII. Future structural and functional studies of these proteins to provide interaction mechanisms will no doubt lead to a better understanding of brain physiology and pathophysiology.

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“…The relative molecular mass of isolated native fodrin molecules has been estimated as 930000 [lo], or close to that predicted if they are tetramers made up of two copies of each subunit. Monoclonal antibody-binding sites [I21 and calm odulin-binding sites [13] mapped on the double-stranded fodrin structure by electron microscopy reveal that each half of the molecule can be envisioned as a heterodimer with two heterodimers bound head-to-head to form the tetramer, similar to erythrocyte spectrin [14].…”

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confidence: 99%

Comparison of spectrin isolated from erythroid and non-erythroid sources

Glenney

1984

Eur J Biochem

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Spectrin from erythrocytes and two other tissues (brain and intestine) were isolated from two distant species, pig and chicken ; some structural and functional properties were compared. A quantitative antibody inhibition assay was used to determine that antibodies to mammalian red cell spectrin cross-react very poorly, if at all, with their nonerythroid (brain) counterpart and similarly antibodies to pig brain spectrin (fodrin) cross-react very weakly with erythroid spectrin. By co rltrast, antibodies which were directed against the 240000-Mr subunit of avian fodrin were completely inhibited with avian spectrin and vice versa. To analyze the structural relatedness of these molecules further we compared the chymotryptic iodinated peptide maps generated from each individual subunit. Consistent with the antibody results, we find little (< 10 %) homology between peptides derived from mammalian fodrin and spectrin, but complete homology (100 %) of the peptides derived from the 240000-M, subunits of chicken fodrin, spectrin and another related molecule from intestine, TW260/240. Whereas the peptide maps of fodrin (brain spectrin) revealed striking similarity between divergent species, suggesting a high degree of structural conservation, the peptide maps of erythrocyte spectrin was highly variable between species, indicating that it has diverged considerably in mammalian evolution. In addition we have compared a functional activity of mammalian spectrins, the ability to bind calm Ddulin, using two different assays. Both results show that, whereas fodrin-calmodulin interaction can be readily demonstrated, the binding to mammalian erythroid spectrin is negligible. This suggests that the high-affinity calmodulin site present on fodrin has been lost from spectrin in mammalian evolution.Within the past few years a number of reports have documented the existence of proteins highly related to red blood cell spectrin in non-erythroid cells [I -61. These molecules, termed fodrin, brain spectrin or calspectrin [2 -4,7,8] isolated from brain tissue, or TW260/240 [2] from brush borders of intestinal epithelial cells, consist of two nonidentical high-molecular-mass subunits and have now been characterized in some detail. They are calmodulin-binding proteins [2,3,9] which bind actin filaments [I01 in a manner similar to erythrocyte spectrin [I I]. Both TW'260/240 [2] and fodrin [2,4,10] have been visualized by low-angle rotary metalshadowing electron microscopy as elongated, flexible doublestranded molecules tightly associated at each end. The relative molecular mass of isolated native fodrin molecules has been estimated as 930000 [lo], or close to that predicted if they are tetramers made up of two copies of each subunit. Monoclonal antibody-binding sites [I21 and calm odulin-binding sites [13] mapped on the double-stranded fodrin structure by electron microscopy reveal that each half of the molecule can be envisioned as a heterodimer with two heterodimers bound head-to-head to form the tetramer, similar to erythrocyte spectrin [...

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Binding sites of calmodulin and actin on the brain spectrin, calspectin.

Cited by 52 publications

References 20 publications

Primary structure of the brain alpha-spectrin [published erratum appears in J Cell Biol 1989 Mar;108(3):following 1175]

Primary structure of the brain alpha-spectrin [published erratum appears in J Cell Biol 1989 Mar;108(3):following 1175]

Brain proteins interacting with the tetramerization region of non-erythroid alpha spectrin

Comparison of spectrin isolated from erythroid and non-erythroid sources

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