ZO-3, a Novel Member of the MAGUK Protein Family Found at the Tight Junction, Interacts with ZO-1 and Occludin
| Autor: | Erika S. Wittchen, Lijie Gu, Bruce R. Stevenson, Jennifer G. Hibbard, Julie Haskins |
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| Rok vydání: | 1998 |
| Předmět: |
DNA
Complementary Protein family Immunoprecipitation Molecular Sequence Data PDZ domain Biology Kidney Zonula Occludens-2 Protein Occludin Cell Line Tight Junctions src Homology Domains Retinoblastoma-like protein 1 Dogs Protein A/G Animals Zonula Occludens Proteins Amino Acid Sequence Cloning Molecular Peptide sequence Base Sequence Sequence Homology Amino Acid Membrane Proteins Cell Biology Phosphoproteins Molecular biology Recombinant Proteins Cingulin Zonula Occludens-1 Protein biology.protein Drosophila Carrier Proteins Sequence Alignment Regular Articles |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| DOI: | 10.1083/jcb.141.1.199 |
| Popis: | A 130-kD protein that coimmunoprecipitates with the tight junction protein ZO-1 was bulk purified from Madin-Darby canine kidney (MDCK) cells and subjected to partial endopeptidase digestion and amino acid sequencing. A resulting 19–amino acid sequence provided the basis for screening canine cDNA libraries. Five overlapping clones contained a single open reading frame of 2,694 bp coding for a protein of 898 amino acids with a predicted molecular mass of 98,414 daltons. Sequence analysis showed that this protein contains three PSD-95/SAP90, discs-large, ZO-1 (PDZ) domains, a src homology (SH3) domain, and a region similar to guanylate kinase, making it homologous to ZO-1, ZO-2, the discs large tumor suppressor gene product of Drosophila, and other members of the MAGUK family of proteins. Like ZO-1 and ZO-2, the novel protein contains a COOH-terminal acidic domain and a basic region between the first and second PDZ domains. Unlike ZO-1 and ZO-2, this protein displays a proline-rich region between PDZ2 and PDZ3 and apparently contains no alternatively spliced domain. MDCK cells stably transfected with an epitope-tagged construct expressed the exogenous polypeptide at an apparent molecular mass of ∼130 kD. Moreover, this protein colocalized with ZO-1 at tight junctions by immunofluorescence and immunoelectron microscopy. In vitro affinity analyses demonstrated that recombinant 130-kD protein directly interacts with ZO-1 and the cytoplasmic domain of occludin, but not with ZO-2. We propose that this protein be named ZO-3. |
| Databáze: | OpenAIRE |
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