C-terminal fragment of amebin promotes actin filament bundling, inhibits acto-myosin ATPase activity and is essential for amoeba migration
| Autor: | Yuriy Rzhepetskyy, Jolanta Jóźwiak, Wanda Kłopocka, Elżbieta Kocik, Magdalena Sobczak, Radosław Skórzewski, Maria Jolanta Redowicz |
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| Rok vydání: | 2010 |
| Předmět: |
Movement
Recombinant Fusion Proteins Molecular Sequence Data Biophysics Protozoan Proteins Arp2/3 complex macromolecular substances In Vitro Techniques Myosins Biochemistry Filamentous actin Microscopy Electron Transmission Myosin Animals Actin-binding protein Amino Acid Sequence Amoeba Molecular Biology Actin biology Actin remodeling Actin cytoskeleton Amoeba proteus biology.organism_classification Peptide Fragments Cell biology Actin Cytoskeleton Multiprotein Complexes biology.protein Rabbits Protein Binding |
| Zdroj: | Archives of biochemistry and biophysics. 506(1) |
| ISSN: | 1096-0384 |
| Popis: | Amebin [formerly termed as ApABP-FI; Sobczak et al. (2007) Biochem. Cell Biol. 85] is encoded in Amoeba proteus by two transcripts, 2672-nt and 1125-nt. A product of the shorter transcript (termed as C-amebin), comprising C-terminal 375 amino-acid-residue fragment of amebin, has been expressed and purified as the recombinant GST-fusion protein. GST-C-amebin bound both to monomeric and filamentous actin. The binding was Ca(2+)-independent and promoted filament bundling, as revealed with the transmission electron microscopy. GST-C-amebin significantly decreased MgATPase activity of rabbit skeletal muscle acto-S1. Removal with endoproteinase ArgC of a positively charged C-terminal region of GST-amebin containing KLASMWEQ sequence abolished actin-binding and bundling as well as the ATPase-inhibitory effect of C-amebin, indicating that this protein region was involved in the interaction with actin. Microinjection of amoebae with antibody against C-terminus of amebin significantly affected amoebae morphology, disturbed cell polarization and transport of cytoplasmic granules as well as blocked migration. These data indicate that amebin may be one of key regulators of the actin-cytoskeleton dynamics and actin-dependent motility in A. proteus. |
| Databáze: | OpenAIRE |
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