کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
6263427 1613896 2014 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Research ReportScreening and identification of dynamin-1 interacting proteins in rat brain synaptosomes
ترجمه فارسی عنوان
گزارش تحقیق: مطالعه و شناسایی پروتئینهای متداول دینامین-1 در سیناپتوزومهای مغز موش صحرایی
کلمات کلیدی
دینین-1، سیناپتوزوم، پروتئین تعامل موش
موضوعات مرتبط
علوم زیستی و بیوفناوری علم عصب شناسی علوم اعصاب (عمومی)
چکیده انگلیسی


- Investigation of interacting proteins of all 4 dynamin-1 functional domains for the first time.
- A total of 63 interacting proteins were identified in this study.
- Fifty-seven interacting proteins were reported here for the first time.
- ClC-3 and Rab GDI interacted with dynamin-1 under native conditions, which was unreported before.

Dynamin-1 is a multi-domain GTPase that is crucial for the fission stage of synaptic vesicle recycling and vesicle trafficking. In this study, we constructed prokaryotic expression plasmids for the four functional domains of dynamin-1, which are pGEX-4T-2-PH, pGEX-4T-2-PRD, pGEX-4T-2-GED and pGEX-4T-2-GTPase. Glutathione S-transferase pull-down, co-immunoprecipitation (co-IP), and liquid chromatography/mass spectrometry were used to screen and identify dynamin-1 interacting proteins in rat brain synaptosomes. We identified a set of 63 candidate protein interactions, including 36 proteins interacting with dynamin-1 C-terminal proline-rich domain (PRD), 14 with pleckstrin-homology domain (PH), 7 with GTPase effector domain (GED) and 6 with GTPase domain, consisting of synaptic vesicle-associated proteins, cytoskeletal proteins, metabolic enzymes and other proteins. We selected three previously unreported dynamin-1 interacting proteins to verify their interaction with dynamin-1 under native conditions. Using co-IP, we found that Rab GDP-dissociation inhibitor (Rab GDI) and chloride channel 3 (ClC-3) do interact with dynamin-1, but not with TUC-4b (the TOAD-64/Ulip/CRMP (TUC) family member). Those novel interactions detected in our study offer valuable insight into the protein-protein interacting network that could enhance our understanding of dynamin-1 mediated synaptic vesicle recycling.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Brain Research - Volume 1543, 16 January 2014, Pages 17-27
نویسندگان
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