Molecular cloning and characterization of DdCAD-1, a Ca2+-dependent cell-cell adhesion molecule, in Dictyostelium discoideum
| Autor: | E F, Wong, S K, Brar, H, Sesaki, C, Yang, C H, Siu |
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| Rok vydání: | 1996 |
| Předmět: |
Myxococcus xanthus
Binding Sites DNA Complementary Base Sequence DNA Plant Sequence Homology Amino Acid Recombinant Fusion Proteins Calcium-Binding Proteins Molecular Sequence Data Cadherins Protein S Rats Animals Calcium Dictyostelium Amino Acid Sequence Cloning Molecular Cell Adhesion Molecules Chickens Glutathione Transferase |
| Zdroj: | The Journal of biological chemistry. 271(27) |
| ISSN: | 0021-9258 4902-2490 |
| Popis: | Dictyostelium discoideum expresses EDTA-sensitive cell-cell adhesion sites soon after the initiation of development, and a Ca2+-binding protein of Mr 24,000 (designated DdCAD-1) has been implicated in this type of adhesiveness. We have previously purified DdCAD-1 to homogeneity and characterized its cell binding activity (Brar, S. K., and Siu, C.-H. (1993) J. Biol. Chem. 268, 24902-24909). In this report, we describe the cloning of DdCAD-1 cDNAs. DNA sequencing revealed a single open reading frame coding for a polypeptide containing 213 amino acids. The identity of the cDNA was confirmed by amino acid sequences of two cyanogen bromide peptides. The deduced amino acid sequence of DdCAD-1 exhibits a relatively high degree of sequence similarity with members of the cadherin family and protein S of Myxococcus xanthus. Unlike the other cadherins, the carboxyl-terminal region of DdCAD-1 contains a Ca2+-binding motif. Although analyses of the sequence suggest that the polypeptide lacks a signal peptide sequence and a transmembrane domain, immunofluorescence microscopy demonstrates the association of DdCAD-1 with the ecto-surface of the plasma membrane. To investigate the structure/function relationships of DdCAD-1, glutathione S-transferase fusion proteins containing different DdCAD-1 fragments were expressed and assayed for their 45Ca2+ and cell binding activities. These studies revealed that the cell binding activity is dependent on the amino-terminal segment and not the carboxyl-terminal Ca2+-binding domain and showed additional Ca2+-binding site(s) within the amino-terminal segment. |
| Databáze: | OpenAIRE |
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