Glycoprotein‐binding site of dystrophin is confined to the cysteine‐rich domain and the first half of the carboxy‐terminal domain

Suzuki, Atsushi; Yoshida, Motoaki; Yamamoto, Hiroyuki; Ozawa, Eijiro

doi:10.1016/0014-5793(92)81265-n

Cited by 199 publications

(126 citation statements)

References 29 publications

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“…The homology of the amino acid sequence between DRP and dystrophin is high for the two domains which have important physiological functions [2,12,24]: (1) the amino-terminal domain which contains the actin-binding site [2,[25][26][27]; and (2) the cysteine-rich and carboxylterminal domains which are involved in the interaction with the sarcolemmal glycoprotein complex in skeletal muscle [28,29]. On the other hand, the sequence homology between these two proteins is relatively low for the rod domain which consists of repeats of triple helix [2,24].…”

Section: Introductionmentioning

confidence: 99%

Purification of dystrophin‐related protein (utrophin) from lung and its identification in pulmonary artery endothelial cells

1993

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Dystrophin-related protein (DRP or utrophin) is an autosomal homologue of dystrophin, a membrane cytoskeletal protein encoded by the Duchenne muscular dystrophy gene. In contrast to dystrophin which is predominantly expressed in muscle, DRP is expressed in various tissues. Here we report the purification and biochemical characterization of DRP from lung which shows the highest levels of DRP expression among adult tissues. DRP was purified from the high alkaline extract of lung membranes using heparin-agarose column chromatography followed by anti-DRP immunoaffinity column chromatography. DRP was expressed in the cultured pulmonary artery endothelial cells. Expression of DRP in endothelial cells could explain its abundant expression in lung. In analogy to dystrophin of muscle cells, DRP could be playing an important role in the mechanical stress mechanisms of endothelial cells.

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Section: Introductionmentioning

confidence: 99%

Purification of dystrophin‐related protein (utrophin) from lung and its identification in pulmonary artery endothelial cells

1993

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“…Dystrophin, the protein responsible for Duchenne muscular dystrophy [ l , 21, is purified as a complex with several proteins called dystrophin-associated proteins (DAPs) from digitonin-solubilized rabbit skeletal muscle membrane [3, 41. At least four DAPs, given the symbols 156DAG, SODAG, 43DAG doublets (A3a and A3b) and 35DAG, were shown to exist in the transmembranous glycoprotein complex (GPC) [5, 61. In the structural study of dystrophin or the complex of dystrophin and its associated proteins (dystrophin-DAP complex) by limited calpain digestion [7], we showed that the locus of GPC binding on the dystrophin molecule is confined to the cysteine-rich domain and the first half of the Cterminal domain [6]. Since this locus is known to be the region whose loss is responsible for Duchenne muscular dystrophy [8], this finding was the first experimental evidence showing that the interaction of GPC with dystrophin is essential to prevent muscle degeneration.…”

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confidence: 99%

Dissociation of the complex of dystrophin and its associated proteins into several unique groups by n‐octyl β‐d‐glucoside

Yoshida

Suzuki

Yamamoto

et al. 1994

European Journal of Biochemistry

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166

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Dystrophin is purified as a complex with several proteins from the digitonin-solubilized muscle cell membrane. Most of dystrophin-associated proteins (DAPs) are assumed to form a large oligomeric transmembranous glycoprotein complex on the sarcolemma and link dystrophin with a basement membrane protein, laminin. In the present study, we found that the purified dystrophin-DAP complex was dissociated into several groups by n-octyl-P-~-glucoside treatment. In particular, we found that the glycoprotein complex stated above was dissociated into two distinct groups: one composed of 156DAG and 43DAG (A3a) and the other composed of SODAG, 35DAG and A3b. We confirmed by crosslinking and immunoaffinity chromatography that these two groups existed in a complexes. We thus concluded that the glycoprotein complex consists of these two subcomplexes. Furthermore, A3b and 43DAG, which had been formerly treated simply as the 43DAG doublets due to their similar electrophoretic mobilities in SDS/PAGE, were shown to be present in two different subcomplexes. Based on the analyses by two-dimensional gel electrophoresis, peptide mapping and immunoblotting, we concluded that A3b is a novel DAP different from 43DAG. Dystrophin, the protein responsible for Duchenne muscular dystrophy [ l , 21, is purified as a complex with several proteins called dystrophin-associated proteins (DAPs) from digitonin-solubilized rabbit skeletal muscle membrane [3, 41. At least four DAPs, given the symbols 156DAG, SODAG, 43DAG doublets (A3a and A3b) and 35DAG, were shown to exist in the transmembranous glycoprotein complex (GPC) [5, 61. In the structural study of dystrophin or the complex of dystrophin and its associated proteins (dystrophin-DAP complex) by limited calpain digestion [7], we showed that the locus of GPC binding on the dystrophin molecule is confined to the cysteine-rich domain and the first half of the Cterminal domain [6]. Since this locus is known to be the region whose loss is responsible for Duchenne muscular dystrophy [8], this finding was the first experimental evidence showing that the interaction of GPC with dystrophin is essential to prevent muscle degeneration.Among the components of GPC, 156DAG [9, 101 and 43DAG [ l l ] were shown to bind directly to laminin and dystrophin, respectively. However, the molecular organization of GPC has not yet been clarified. Thus, its study is indispensable in order to clarify the molecular basis behind muscular dystrophy. We showed immunochemically that the components of GPC (50DAG and 35DAG) are expressed in striated muscles but not in smooth muscles such as uterus and aorta, whereas 43DAG is rather ubiquitously expressed in various tissues [12, 131. On the other hand, it was reported that in the muscles of patients with severe childhood autosoma1 recessive muscular dystrophy (SCARMD), 5ODAG is lost and 35DAG is reduced, while dystrophin and other DAPs are preserved [ 141. Similar observations were made in the skeletal muscle of dystrophic hamster [15, 161. We also observed that in the skeletal...

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“…While the precise function of dystrophin is unknown, its absence leads to several pathophysiological changes that ultimately result in profound necrosis. Biochemical studies have led to the identification of a dystrophin-glycoprotein complex (DGC) that links the sarcolemma and extracellular matrix to the carboxy terminus of dystrophin [9,10,11,12]. Interestingly, this region of dystrophin contains two leucine zipper motifs separated by a 44 amino acid proline rich spacer [2].…”

Section: Introductionmentioning

confidence: 99%

Troponin T is capable of binding dystrophin via a leucine zipper

Pearlman¹,

Powaser²,

Elledge³

et al. 1994

FEBS Letters

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Using genetic and physical assays for protein-protein interactions, we identified a fast isoform of troponin T that binds to dystrophin. Troponin T specifically bound to the first of two highly conserved leucine zipper motifs in the carboxy terminus of dystrophin [1,2]. Single amino acid changes in the zipper predicted to disrupt a-helix formation or cause steric hindrance abolished this binding. These data support the hypothesis that dystrophin couples the contractile apparatus to the sarcolemma and indicate that leucine zipper mediated protein-protein interactions are functionally important in the cytoskeleton as well as the nucleus.

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Glycoprotein‐binding site of dystrophin is confined to the cysteine‐rich domain and the first half of the carboxy‐terminal domain

Cited by 199 publications

References 29 publications

Purification of dystrophin‐related protein (utrophin) from lung and its identification in pulmonary artery endothelial cells

Purification of dystrophin‐related protein (utrophin) from lung and its identification in pulmonary artery endothelial cells

Dissociation of the complex of dystrophin and its associated proteins into several unique groups by n‐octyl β‐d‐glucoside

Troponin T is capable of binding dystrophin via a leucine zipper

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