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Title: Structural characterization of the self‐association domain of swallow
Abstract

Swallow, a 62 kDa multidomain protein, is required for the proper localization of several mRNAs involved in the development of Drosophila oocytes. The dimerization of Swallow depends on a 71‐residue self‐association domain in the center of the protein sequence, and is significantly stabilized by a binding interaction with dynein light chain (LC8). Here, we detail the use of solution‐state nuclear magnetic resonance spectroscopy to characterize the structure of this self‐association domain, thereby establishing that this domain forms a parallel coiled‐coil and providing insight into how the stability of the dimerization interaction is regulated.

 
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PAR ID:
10364286
Author(s) / Creator(s):
 ;  
Publisher / Repository:
Wiley Blackwell (John Wiley & Sons)
Date Published:
Journal Name:
Protein Science
Volume:
30
Issue:
5
ISSN:
0961-8368
Page Range / eLocation ID:
p. 1056-1063
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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    This article was corrected on 25 July 2022. See the end of the full text for details.

    Basic Protocol 1: Genetic engineering strategy for the generation of modular light‐activated protein dimerization units

    Support Protocol 1: Molecular cloning

    Basic Protocol 2: Cell culture and transfection

    Support Protocol 2: Production of dark containers for optogenetic samples

    Basic Protocol 3: Confocal microscopy and light‐dependent activation of the dimerization system

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    Alternate Protocol 2: Induction of organelles’ membrane tethering

    Alternate Protocol 3: Optogenetic reconstitution of protein function

    Basic Protocol 4: Image analysis

    Support Protocol 3: Analysis of apparent on‐ and off‐kinetics

    Support Protocol 4: Analysis of changes in organelle overlap over time

     
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