Journal:Acta Cryst F:S2053230X20016015
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<b>Molecular Tour</b><br> | <b>Molecular Tour</b><br> | ||
| - | We were unable to express the wild-type kinase domain (1-346) of hematopoietic progenitor kinase 1 (HPK1, also called MAP4K1). We investigated the potential surface aggregation properties of a model of HPK1 and chose eight sites to investigate by yeast surface display. This led to a double mutant L221D F225E, which expressed in baculovirus-infected insect cells at multiple milligrams per liter of culture. Purification of this construct showed that it was truncated after residue 319, which led to a revised construct of HPK1(1-319) L221D F225E. This construct crystallized in the presence of inhibitors and led to a structure with two molecules/asymmetric unit. The activation loop on both molecules was domain-swapped, that is the activation loop of chain A bind to chain B and vice versa. We were able to use this construct to determine many structures of HPK1/inhibitor complexes in support a structure-based drug design effort. | + | We were unable to express the wild-type kinase domain (1-346) of hematopoietic progenitor kinase 1 (HPK1, also called MAP4K1). We investigated the potential surface aggregation properties of a model of HPK1 and chose eight sites to investigate by yeast surface display. This led to a double mutant L221D F225E, which expressed in baculovirus-infected insect cells at multiple milligrams per liter of culture. Purification of this construct showed that it was truncated after residue 319, which led to a revised construct of HPK1(1-319) L221D F225E. This construct crystallized in the presence of inhibitors and led to a structure with two molecules/asymmetric unit. The activation loop on both molecules was domain-swapped, that is the activation loop of chain A bind to chain B and ''vice versa''. We were able to use this construct to determine many structures of HPK1/inhibitor complexes in support a structure-based drug design effort. |
<scene name='87/870564/Cv/4'>The contents of the asymmetric unit</scene> are shown with chain A in cyan and chain B in violet. Between the two monomers one can see the domain swapping of the activation loop. <scene name='87/870564/Cv/6'>Superposition of the two monomers</scene> (chain A in cyan and chain B in violet) showing that with the exception of the position of the αC-helix and surrounding residues, the activation loop and the extended C-terminus of chain A, the chains follow the same path. | <scene name='87/870564/Cv/4'>The contents of the asymmetric unit</scene> are shown with chain A in cyan and chain B in violet. Between the two monomers one can see the domain swapping of the activation loop. <scene name='87/870564/Cv/6'>Superposition of the two monomers</scene> (chain A in cyan and chain B in violet) showing that with the exception of the position of the αC-helix and surrounding residues, the activation loop and the extended C-terminus of chain A, the chains follow the same path. | ||
Revision as of 15:20, 22 December 2020
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This page complements a publication in scientific journals and is one of the Proteopedia's Interactive 3D Complement pages. For aditional details please see I3DC.
