دورية أكاديمية
Phosphorylation mutants elucidate the mechanism of annexin IV-mediated membrane aggregation
| العنوان: | Phosphorylation mutants elucidate the mechanism of annexin IV-mediated membrane aggregation |
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| المؤلفون: | Kaetzel, M A, Mo, Y D, Mealy, T R, Campos, B, Bergsma-Schutter, W, Brisson, A, Dedman, J R, Seaton, B A |
| المصدر: | Kaetzel , M A , Mo , Y D , Mealy , T R , Campos , B , Bergsma-Schutter , W , Brisson , A , Dedman , J R & Seaton , B A 2001 , ' Phosphorylation mutants elucidate the mechanism of annexin IV-mediated membrane aggregation ' , Biochemistry , vol. 40 , no. 13 , pp. 4192-4199 . https://doi.org/10.1021/bi002507sTest |
| سنة النشر: | 2001 |
| المجموعة: | University of Groningen research database |
| مصطلحات موضوعية: | N-TERMINAL DOMAIN, ACTIVATED CHLORIDE CONDUCTANCE, CALCIUM-DEPENDENT BINDING, PHOSPHOLIPID-BINDING, VESICLE AGGREGATION, ELECTRON-MICROSCOPY, LIPOSOME AGGREGATION, PROTEIN, CRYSTALLOGRAPHY, CRYSTALLIZATION |
| الوصف: | Site-directed mutagenesis, electron microscopy, and X-ray crystallography were used to probe the structural basis of annexin IV-induced membrane aggregation and the inhibition of this property by protein kinase C phosphorylation. Site-directed mutants that either mimic (Thr6Asp, T6D) or prevent (Thr6Ala, T6A) phosphorylation of threonine 6 were produced for these studies and compared with wildtype annexin IV. In vitro assays showed that unmodified wild-type annexin IV and the T6A mutant, but not PKC-phosphorylated wild-type or the T6D mutant, promote vesicle aggregation. Electron crystallographic data of wild-type and T6D annexin IV revealed that, similar to annexin V, the annexin IV proteins form 2D trimer-based ordered arrays on phospholipid monolayers. Cryo-electron microscopic images of junctions formed between lipid vesicles in the presence of wild-type annexin IV indicated a separation distance corresponding to the thickness of two layers of membrane-bound annexin IV. In this orientation, a single layer of WT annexin IV, attached to the outer leaflet of one vesicle, would undergo face-to-face self-association with the annexin layer of a second vesicle. The 2.0-Angstrom resolution crystal structure of the T6D mutant showed that the mutation causes release of the N-terminal tail from the protein core. This change would preclude the face to-face annexin self-association required to aggregate vesicles. The data suggest that reversible complex formation through phosphorylation and dephosphorylation could occur in vivo and play a role in the regulation of vesicle trafficking following changes in physiological states. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| العلاقة: | https://research.rug.nl/en/publications/57f284d2-c10c-46bf-8525-a60585b637ebTest |
| DOI: | 10.1021/bi002507s |
| الإتاحة: | https://doi.org/10.1021/bi002507sTest https://hdl.handle.net/11370/57f284d2-c10c-46bf-8525-a60585b637ebTest https://research.rug.nl/en/publications/57f284d2-c10c-46bf-8525-a60585b637ebTest |
| حقوق: | info:eu-repo/semantics/closedAccess |
| رقم الانضمام: | edsbas.7A58F7E5 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1021/bi002507s |
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