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Updated: Mar 30, 2026

Chemical Modification of the Tryptophan Residue in a Recombinant Ca2+-ATPase N-domain for Studying Tryptophan-ANS FRET
Published on: October 9, 2021
Structural basis for the binding of tryptophan-based motifs by δ-COP
Richard J Suckling1, Pak Phi Poon2, Sophie M Travis3
1Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom; Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, United Kingdom;
Coatomer
Area of Science:
- Cell biology
- Structural biology
- Protein-protein interactions
Background:
- Coatomer is essential for COPI-coated vesicle formation.
- It comprises two subcomplexes: F-subcomplex (Arf1:GTP, βγδζ-COP) and B-subcomplex (αβ'ε-COP).
- The F-subcomplex targets coatomer to membranes, while the B-subcomplex binds cargo.
Purpose of the Study:
- To elucidate the structural basis of δ-COP subunit interactions with binding partners.
- To investigate the role of tryptophan-based motifs in coatomer function.
- To understand how these interactions influence COPI vesicle trafficking.
Main Methods:
- X-ray crystallography to determine the structure of yeast δ-COP micro-homology domain with a WxW motif.
- Isothermal titration calorimetry (ITC) to confirm binding modes.
- Site-directed mutagenesis to assess the functional impact of mutations.
Main Results:
- The structure reveals that yeast δ-COP binds a WxW motif from Dsl1 tether at a distinct site from AP adaptors.
- Saccharomyces cerevisiae Gcs1p uses a WxxF motif to bind δ-COP similarly.
- Mammalian δ-COP also binds related tryptophan-based motifs, such as from ArfGAP1.
Conclusions:
- δ-COP subunits bind Wxn(1-6)[WF] motifs in unstructured protein regions.
- These interactions are crucial for the lifecycle of COPI-coated vesicles.
- Disruption of the δ-COP binding site in yeast leads to growth and protein trafficking defects.
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