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Characterization of the binding interaction between the oncoprotein gankyrin and a grafted S6 ATPase.

Chapman AM, Rogers BE, McNaughton BR - Biochemistry (2014)

Bottom Line: A complex with the C-terminal portion of the proteosomal subunit S6 ATPase is the only available structure of a protein-protein interaction involving the oncoprotein gankyrin.However, difficulties associated with recombinant expression of S6 ATPase alone, or truncations thereof, have limited our understanding of this assembly.We replaced the C-terminal portion of FtsH from Escherichia coli with the structurally homologous C-terminal portion of S6 ATPase and used this grafted protein to characterize the gankyrin-S6 ATPase binding interaction by isothermal titration calorimetry.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and ‡Department of Biochemistry & Molecular Biology, Colorado State University , Fort Collins, Colorado 80523, United States.

ABSTRACT
A complex with the C-terminal portion of the proteosomal subunit S6 ATPase is the only available structure of a protein-protein interaction involving the oncoprotein gankyrin. However, difficulties associated with recombinant expression of S6 ATPase alone, or truncations thereof, have limited our understanding of this assembly. We replaced the C-terminal portion of FtsH from Escherichia coli with the structurally homologous C-terminal portion of S6 ATPase and used this grafted protein to characterize the gankyrin-S6 ATPase binding interaction by isothermal titration calorimetry.

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(a) Complex between gankyrin (orange) and the C-terminal portionof the S6 ATPase subunit of the 26S proteosome [red, Protein DataBank (PDB) entry 2DVW]. The direction of the arrow next to each protein indicates thedirection of a 90° rotation, which reveals the binding surfaces,as shown in panel b. (b) Gankyrin and S6 ATPase binding face residuescritical to complex stabilization (and mutated in this work). (c)S6 ATPase superimposed on the C-terminal domain of FtsH (PDB entry ILV7).
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fig1: (a) Complex between gankyrin (orange) and the C-terminal portionof the S6 ATPase subunit of the 26S proteosome [red, Protein DataBank (PDB) entry 2DVW]. The direction of the arrow next to each protein indicates thedirection of a 90° rotation, which reveals the binding surfaces,as shown in panel b. (b) Gankyrin and S6 ATPase binding face residuescritical to complex stabilization (and mutated in this work). (c)S6 ATPase superimposed on the C-terminal domain of FtsH (PDB entry ILV7).

Mentions: Overexpression of gankyrin (Figure 1a, orange)is directly linked to the onset, proliferation,and/or metastasis of breast,1,2 liver,3 oral,4 pancreatic,5 and colorectal cancers.6 In addition, gankyrin plays an essential role in Ras-initiated tumorigenesis,which is operative in ∼30% of all cancers.7 Protein–protein interactions (PPIs) involving gankyrinare of great interest in basic research and as therapeutic targets.


Characterization of the binding interaction between the oncoprotein gankyrin and a grafted S6 ATPase.

Chapman AM, Rogers BE, McNaughton BR - Biochemistry (2014)

(a) Complex between gankyrin (orange) and the C-terminal portionof the S6 ATPase subunit of the 26S proteosome [red, Protein DataBank (PDB) entry 2DVW]. The direction of the arrow next to each protein indicates thedirection of a 90° rotation, which reveals the binding surfaces,as shown in panel b. (b) Gankyrin and S6 ATPase binding face residuescritical to complex stabilization (and mutated in this work). (c)S6 ATPase superimposed on the C-terminal domain of FtsH (PDB entry ILV7).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230329&req=5

fig1: (a) Complex between gankyrin (orange) and the C-terminal portionof the S6 ATPase subunit of the 26S proteosome [red, Protein DataBank (PDB) entry 2DVW]. The direction of the arrow next to each protein indicates thedirection of a 90° rotation, which reveals the binding surfaces,as shown in panel b. (b) Gankyrin and S6 ATPase binding face residuescritical to complex stabilization (and mutated in this work). (c)S6 ATPase superimposed on the C-terminal domain of FtsH (PDB entry ILV7).
Mentions: Overexpression of gankyrin (Figure 1a, orange)is directly linked to the onset, proliferation,and/or metastasis of breast,1,2 liver,3 oral,4 pancreatic,5 and colorectal cancers.6 In addition, gankyrin plays an essential role in Ras-initiated tumorigenesis,which is operative in ∼30% of all cancers.7 Protein–protein interactions (PPIs) involving gankyrinare of great interest in basic research and as therapeutic targets.

Bottom Line: A complex with the C-terminal portion of the proteosomal subunit S6 ATPase is the only available structure of a protein-protein interaction involving the oncoprotein gankyrin.However, difficulties associated with recombinant expression of S6 ATPase alone, or truncations thereof, have limited our understanding of this assembly.We replaced the C-terminal portion of FtsH from Escherichia coli with the structurally homologous C-terminal portion of S6 ATPase and used this grafted protein to characterize the gankyrin-S6 ATPase binding interaction by isothermal titration calorimetry.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and ‡Department of Biochemistry & Molecular Biology, Colorado State University , Fort Collins, Colorado 80523, United States.

ABSTRACT
A complex with the C-terminal portion of the proteosomal subunit S6 ATPase is the only available structure of a protein-protein interaction involving the oncoprotein gankyrin. However, difficulties associated with recombinant expression of S6 ATPase alone, or truncations thereof, have limited our understanding of this assembly. We replaced the C-terminal portion of FtsH from Escherichia coli with the structurally homologous C-terminal portion of S6 ATPase and used this grafted protein to characterize the gankyrin-S6 ATPase binding interaction by isothermal titration calorimetry.

Show MeSH
Related in: MedlinePlus