Reference - Detail
| RRC ID | 28934 |
|---|---|
| Author | Eichinger L, Schleicher M. |
| Title | Characterization of actin- and lipid-binding domains in severin, a Ca(2+)-dependent F-actin fragmenting protein. |
| Journal | Biochemistry |
| Abstract |
Severin is a Ca(2+)-activated actin-binding protein that nucleates actin assembly and severs and caps the fast growing ends of actin filaments. It consists of three highly conserved domains. To investigate the domain structure of severin, we constructed genetically the N-terminal domain 1, the middle domain 2, and the tandem domains 2 + 3. Their interaction with actin, Ca2+, and lipids was characterized. Domain 1 contains the F-actin capping and a Ca(2+)-binding site [Eichinger, L., Noegel, A. A., & Schleicher, M. (1991) J. Cell Biol. 112, 665-676]. Binding of domain 2 to actin filaments was Ca(2+)-dependent and saturated at a 1:1 molar ratio. In the presence of Ca2+, about 1.5 mol of domains 2 + 3 bound per mole of F-actin subunit. Scatchard analysis gave a Kd of 18 microM for the interaction of domain 2 with F-actin subunits and a Kd of 1.6 microM for domains 2 + 3. Low-shear viscometry, electron microscopy, and low-speed sedimentation assays showed that domains 2 + 3 induced bundling of actin filaments. The influence of PIP2 micelles on the different activities of severin was assayed using native severin and N- and C-terminally truncated fragments. Severin contains at least two PIP2-binding sites since the activities of the two nonoverlapping severin fragments domain 1 and domains 2 + 3 were inhibited by PIP2. The specificity of severin-phospholipid interaction was investigated by studying the regulation of native severin by PIP2 and other pure or mixed phospholipids.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Volume | 31(20) |
| Pages | 4779-87 |
| Published | 1992-5-26 |
| DOI | 10.1021/bi00135a006 |
| PMID | 1591239 |
| MeSH | Actins / chemistry* Actins / metabolism Animals Base Sequence Calcium-Binding Proteins / chemistry* Carrier Proteins / chemistry* Carrier Proteins / isolation & purification Fungal Proteins / antagonists & inhibitors Fungal Proteins / chemistry* Microfilament Proteins / antagonists & inhibitors Microfilament Proteins / chemistry* Microfilament Proteins / isolation & purification Molecular Sequence Data Peptide Fragments / chemistry Peptide Fragments / drug effects* Phospholipids / chemistry* Phospholipids / pharmacology Protein Conformation Protozoan Proteins* Rabbits Structure-Activity Relationship |
| IF | 2.865 |
| Times Cited | 54 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Cellular slime molds | S00412 |