Lipid-binding role of betaII-spectrin ankyrin-binding domain
| Autor: | Aleksander F. Sikorski, Patrycja M. Dubielecka, Ewa Plażuk, Anna Chorzalska, Witold Diakowski, Agnieszka Szmaj, Marek Langner, Ewa Bok, Katarzyna Stebelska, Marek Lisowski, Anita Hryniewicz-Jankowska |
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| Rok vydání: | 2007 |
| Předmět: |
Ankyrins
Protein Folding animal structures Recombinant Fusion Proteins Green Fluorescent Proteins Molecular Sequence Data Melanoma Experimental Ankyrin binding Biology Cell Line Membrane Lipids Mice Structure-Activity Relationship Protein structure Protein Interaction Mapping Escherichia coli Ankyrin Animals Humans Spectrin Binding site Sequence Deletion chemistry.chemical_classification Binding Sites Vesicle Circular Dichroism fungi Microfilament Proteins EPB41 Cell Biology General Medicine Transfection Surface Plasmon Resonance Protein Structure Tertiary chemistry Biochemistry Microscopy Fluorescence Liposomes Biophysics Mutagenesis Site-Directed Carrier Proteins Sequence Alignment Algorithms HeLa Cells |
| Zdroj: | Cell biology international. 31(12) |
| ISSN: | 1065-6995 |
| Popis: | It is known that erythroid and non-erythroid spectrins binding of vesicles and monolayers containing PE proved sensitive to inhibition by red blood cell ankyrin. We now show that the bacterially-expressed recombinant peptides representing betaII(brain)-spectrin's ankyrin-binding domain and its truncated mutants showed lipid-binding activity, although only those containing a full-length amino terminal fragment showed high to moderate affinity towards phospholipid mono- and bilayers and a substantial sensitivity of this binding to inhibition by ankyrin. These results are in accordance with our published data on betaI-spectrin's ankyrin-binding domain [Hryniewicz-Jankowska A, et al. Mapping of ankyrin-sensitive, PE/PC mono- and bilayer binding site in erythroid beta-spectrin. Biochem J 2004;382:677-85]. Moreover, we tested also the effect of transient transfection of living cells of several cell-lines with vectors coding for GFP-conjugates including betaII and also betaI full-length ankyrin-binding domain and their truncated fragments on the membrane skeleton organization. The transfection with constructs encoding full-length ankyrin-binding domain of betaII and betaI spectrin resulted in increased aggregation of membrane skeleton and its punctate appearance in contrast to near normal appearance of membrane skeleton of cells transiently transfected with GFP control or construct encoding ankyrin-binding domain truncated at their N-terminal region. Our results therefore indicate the importance of N-terminal region for lipid-binding activity of the beta-spectrin ankyrin-binding domain and its substantial role in maintaining the spectrin-based skeleton distribution. |
| Databáze: | OpenAIRE |
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