Molecular organization of the desmoglein-plakoglobin complex
| Autor: | Regina B. Troyanovsky, Sergey M. Troyanovsky, Nikolai A. Chitaev, Alexander Z. Averbakh |
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| Rok vydání: | 1998 |
| Předmět: |
Molecular Sequence Data
Plakoglobin Desmocollins Biology Transfection Desmoglein Cell Line Desmosome medicine Humans Amino Acid Sequence Desmosomal Cadherins Alanine Cadherin Cortical actin cytoskeleton Epithelial Cells Desmosomes Cell Biology Alanine scanning Recombinant Proteins Cell biology Cytoskeletal Proteins Kinetics Intercellular Junctions medicine.anatomical_structure Desmoplakins Biochemistry Mutagenesis Site-Directed gamma Catenin Desmogleins Cell Adhesion Molecules |
| Zdroj: | Journal of Cell Science. 111:1941-1949 |
| ISSN: | 1477-9137 0021-9533 |
| DOI: | 10.1242/jcs.111.14.1941 |
| Popis: | Different epithelial intercellular junctions contain distinct complexes incorporating plakoglobin. In adherens junctions, plakoglobin interacts with two molecules, the transmembrane adhesion protein of the cadherin family (e.g. E-cadherin) and alpha-catenin. The latter is thought to anchor the cadherin-plakoglobin complex to the cortical actin cytoskeleton. In desmosomes, plakoglobin forms a complex with desmosomal cadherins, either desmoglein (Dsg) or desmocollin (Dsc), but not with alpha-catenin. To further understand the structure and assembly of the plakoglobin-cadherin complexes we analyzed amino acid residues involved in plakoglobin-Dsg interactions using alanine scanning mutagenesis. Previously, we have shown that plakoglobin interacts with a 72 amino acid-long cytoplasmic domain (C-domain) that is conserved among desmosomal and classic cadherins. In this paper, we show that a row of the large hydrophobic residues located at the C-terminal portion of the Dsg C-domain is indispensable for interaction with plakoglobin. To study a reciprocal site we expressed plakoglobin (MPg) or its mutants tagged by 6 myc epitope in epithelial A-431 cells. Using sucrose gradient centrifugation and subsequent co-immunoprecipitation, MPg was found to be efficiently incorporated into the same type of complexes as endogenous plakoglobin. A major pool of Dsg-plakoglobin complexes sedimented at 8S and exhibited a 1:1 stoichiometry. Using alanine scanning mutagenesis and the co-immunoprecipitation assay we identified nine hydrophobic amino acids within the arm repeats 1–3 of plakoglobin, that are required for binding to Dsg and Dsc. Eight of these amino acids also participate in the interaction with alpha-catenin. No mutations were found to reduce the affinity of plakoglobin binding to E-cadherin. These data provide direct evidence that the same hydrophobic plakoglobin surface is essential for mutually exclusive interaction with distinct proteins such as alpha-catenin and desmosomal cadherins. |
| Databáze: | OpenAIRE |
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