Clarissa G. Jakob

Here we describe the three-dimensional crystal structures of human glucocorticoid receptor ligand-binding domain (GR-LBD) in complex with the antagonist RU-486 at 2.3 Å resolution and with the agonist dexamethasone ligand together with a coactivator peptide at 2.8 Å. The RU-486 structure was solved in several different crystal forms, two with helix 12 intact (GR1 and GR3) and one with a protease-digested C terminus (GR2). In GR1, part of helix 12 is in a position that covers the co-activator pocket, whereas in the GR3, domain swapping is seen between the crystallographically identical subunits in the GR dimer. An arm consisting of the end of helix 11 and beyond stretches out from one molecule, and helix 12 binds to the other LBD, partly blocking the coactivator pocket of that molecule. This type of GR-LBD dimer has not from the subunits of the a between the in the two RU-486 GR-LBD structures that GR has a between the end of helix 11 and the end of helix Here we describe the three-dimensional crystal structures of human glucocorticoid receptor ligand-binding domain (GR-LBD) in complex with the antagonist RU-486 at 2.3 Å resolution and with the agonist dexamethasone ligand together with a coactivator peptide at 2.8 Å. The RU-486 structure was solved in several different crystal forms, two with helix 12 intact (GR1 and GR3) and one with a protease-digested C terminus (GR2). In GR1, part of helix 12 is in a position that covers the co-activator pocket, whereas in the GR3, domain swapping is seen between the crystallographically identical subunits in the GR dimer. An arm consisting of the end of helix 11 and beyond stretches out from one molecule, and helix 12 binds to the other LBD, partly blocking the coactivator pocket of that molecule. This type of GR-LBD dimer has not from the subunits of the a between the in the two RU-486 GR-LBD structures that GR has a between the end of helix 11 and the end of helix The glucocorticoid receptor and glucocorticoid LBD, ligand-binding and glucocorticoid LBD, ligand-binding is a of the receptor part of the of the of receptor and has of glucocorticoid receptor The structure of the receptor was from of GR The of a receptor to a the glucocorticoid was GR and GR was the receptor to The three-dimensional structure of a receptor was the structure of the GR domain was solved the structure of the GR ligand-binding domain has The structures of other receptor of of the of and the GR-LBD structure was in complex with the agonist dexamethasone and a coactivator peptide the and to with the and is in ligand and is a glucocorticoid and and the with to GR and the is a GR and to the of the and is a of of of the and a the of the with other of the receptor the of of the GR-LBD and with the we out a of of the GR-LBD This of the of and with a structure the In we describe the structure of in with the antagonist RU-486 in with the agonist RU-486 is and that has in is a The of the of RU-486 has to and not the blocking of agonist the of GR binds to with a The antagonist to a that the with is that is the ligand of has with to of dexamethasone and RU-486 of dexamethasone not RU-486 of This was to the of the of of RU-486 the of at the the of of receptor and receptor This is the domain of the is complex and the ligand-binding domain In RU-486 agonist in the of the The agonist of RU-486 the of part of the of in of RU-486 has with a of the between the structures of GR to dexamethasone with RU-486 is of to the ligand different one receptor helix from the receptor structures and different the with the of a GR that has a ligand-binding pocket and is consisting of was at the the C different to the and GR was in and The was with the of was The with the human GR-LBD The and in a at the was in and at from of in the with a at The was and was to The was to to the The GR-LBD was with and The was from at The was a and with a The was at and a to the GR-LBD RU-486 the was at and GR-LBD was to in a to the in of of and The at 12 12 and with of coactivator peptide from and and at the at and the at the at and and with the in whereas and and The structure was solved in a GR from a The structures solved the GR structure at the and crystal The structure was solved in the crystal to Å resolution of and in crystal was not a was that to 2.8 and the was (GR2). In structure the helix 12 was of that to 2.3 Å and with intact C the of The of GR-LBD in and to Å resolution The in The the two crystal forms, and of and of GR in seen in from of in a The the has to GR The in the The to a of crystal was the with and to a of the of coactivator peptide to GR-LBD with The crystal to and the the crystal a and crystal GR3, the one to was solved in a of GR helix in the in the the that was with of and a of to the in was in The the of and to RU-486 in the the the part of the structure was with the in from a and in the The structure was to with and has in and The In the of and in a the agonist structure a to was in with a the structure and the of the structure from the of helix and to with in the was consisting of a dimer and a in the The not the molecule. This with a of the of the of of the from a molecule. with of structure from the the the of a of was out and at of and of a with the This with the that the in the the and with a of the the of the to and the of to and the and and to at the terminus in of the in the The structure is of with and a in crystal is in GR-LBD structures of of in The structures of and GR to of other in to the receptor and with a of Å and with of Å and 12 not The structures a In GR3, the of helix 12 from the agonist position the ligand-binding pocket in the seen with to the of In the seen of the from of the receptor the RU-486 is one helix 12 in in the in at 2.3 and 2.8 Å resolution of antagonist and the is and a molecule, in to the of dexamethasone is in from The is in with The of GR the and the agonist structures to antagonist from agonist one is of the of helix 12 that in the structure In that of antagonist to the in of helix 12 that of the of RU-486 of helix 12 in the agonist position is seen in the In GR3, helix 11 is at whereas in the helix 11 to of a the structure is out and the is Å and the out the from to a The is with the in other that of the In GR3, the of is to the of the RU-486 is between the two the the between 11 and 12 the other of the dimer and binds in a between the agonist position of helix 12 and the coactivator The of helix 12 in is to that of the position that of helix 12 of the and structures that the helix 12 is to the of the helix 12 position of and the pocket that is has in the of a at the end of helix 11 was is is in the at has the that is from in with of of that and that the of the ligand in and the not GR a of was seen the of between and This of the position of the C terminus of GR in the agonist of GR with dexamethasone and with that seen in is a Å between the helix 12 and the in the part This to that helix 12 the and with C terminus to molecule. This to in the GR with In the of structures with the helix 12 of in the The RU-486 is in the other in The is in that the from the in the of the to and and a the the other the a to and a to the of has to is a that a position in the and antagonist and agonist a different is In the agonist structure is with the ligand and a Å to to position in the In the RU-486 from the ligand and with that in the structure helix 12 a position the ligand-binding pocket, the peptide to in the co-activator a in and the of RU-486 helix 12 from the agonist position in helix 12 is and covers the co-activator the receptor from a is a that has with the of In with GR we the with to in the the the GR to and the was not the was the in the of was to in of in of the was that the of the with a that was to the of GR was This is stretches a In is a of the other pocket between and the GR structure from and structures the type at of with that the a of in the of is in the and a to In the type in and with the of the The in from the structure with the In the type and the a to In the the other out from the pocket and The is and one molecule, of is solved to resolution and not the pocket not in the the and in the The of not to seen a of and of the of a of and 12 in is the coactivator peptide in is The in the structure in and The of not the of GR seen in In was of the the one is a the in the ligand and binds to the agonist structure was solved in in to the structure that was The in the that we two and The dimer in is the to the GR dimer from In GR the of the GR dimer from structure to that GR in the intact of the of the In that of the The the is not in the In the at that the is the and with ligand is in the of the ligand of the agonist structures and that the of the is in the with and at identical the of the in and the the other between different with between in the part of the ligand-binding The in structure between the receptor is seen in in In a to the The position of the of the other in GR-LBD that to the of the between the to a in of GR and and different structures of GR with the antagonist structure helix 12 the part of the coactivator pocket This with is seen in other receptor antagonist In the is 12 together with the end of helix 11 and the out to the agonist and coactivator of a The part of the between 11 and 12 a helix helix the to the position seen in the structure in the agonist structure 12 is the to GR and the of helix The structure is the of the structures together of with other in of different The structures that several to 12 different the is in the 12 is and the coactivator peptide is helix 12 in binds in a position to that of with a of helix 12 and a coactivator peptide in in complex with The glucocorticoid receptor and glucocorticoid LBD, ligand-binding and glucocorticoid LBD, ligand-binding is a of the receptor part of the of the of receptor and has of glucocorticoid receptor The structure of the receptor was from of GR The of a receptor to a the glucocorticoid was GR and GR was the receptor to The three-dimensional structure of a receptor was the structure of the GR domain was solved the structure of the GR ligand-binding domain has The structures of other receptor of of the of and the GR-LBD structure was in complex with the agonist dexamethasone and a coactivator peptide The the and to with the and is in ligand and is a glucocorticoid and and the with to GR and the is a GR and to the of the and is a of of of the and a the of the with other of the receptor the of of the GR-LBD and with the we out a of of the GR-LBD This of the of and with a structure the In we describe the structure of in with the antagonist RU-486 in with the agonist The RU-486 is and that has in is a The of the of RU-486 has to and not the blocking of agonist the of GR binds to with a The antagonist to a that the with is that is the ligand of has with to of dexamethasone and RU-486 of dexamethasone not RU-486 of This was to the of the of of RU-486 the of at the the of of receptor and receptor This is the domain of the is complex and the ligand-binding domain In RU-486 agonist in the of the The agonist of RU-486 the of part of the of in of RU-486 has with a of the between the structures of GR to dexamethasone with RU-486 is of to the ligand different one receptor helix from the receptor structures and different the with the of a GR that has a ligand-binding pocket and is consisting of was at the the C different to the and GR was in and The was with the of was The with the human GR-LBD The and in a at the was in and at from of in the with a at The was and was to The was to to the The GR-LBD was with and The was from at The was a and with a The was at and a to the GR-LBD RU-486 the was at and GR-LBD was to in a to the in of of and The at 12 12 and with of coactivator peptide from and and at the at and the at the at and and with the in whereas and and The structure was solved in a GR from a The structures solved the GR structure at the and different the with the of a GR that has a ligand-binding pocket and is consisting of was at the the C different to the and GR was in and The was with the of was The with the human GR-LBD The and in a at the was in and at from of in the with a at The was and was to The was to to the The GR-LBD was with and The was from at The was a and with a The was at and a to the GR-LBD RU-486 the was at and GR-LBD was to in a to the in of of and The at 12 12 and with of coactivator peptide from and and at the at and the at the at and and with the in whereas and and The structure was solved in a GR from a The structures solved the GR structure at the and crystal The structure was solved in the crystal to Å resolution of and in crystal was not a was that to 2.8 and the was (GR2). In structure the helix 12 was of that to 2.3 Å and with intact C the of The of GR-LBD in and to Å resolution The in The the two crystal forms, and of and of GR in seen in from of in a The the has to GR The in the The to a of crystal was the with and to a of the of coactivator peptide to GR-LBD with The crystal to and the the crystal a and crystal GR3, the one to was solved in a of GR helix in the in the the that was with of and a of to the in was in The the of and to RU-486 in the the the part of the structure was with the in from a and in the The structure was to with and has in and The In the of and in a the agonist structure a to was in with a the structure and the of the structure from the of helix and to with in the was consisting of a dimer and a in the The not the molecule. This with a of the of the of of the from a molecule. with of structure from the the the of a of was out and at of and of a with the This with the that the in the the and with a of the the of the to and the of to and the and and to at the terminus in of the in the The structure is of with and a in crystal is in GR-LBD structures of of in The structures of and GR to of other in to the receptor and with a of Å and with of Å and 12 not The structures a In GR3, the of helix 12 from the agonist position the ligand-binding pocket in the seen with to the of In the seen of the from of the receptor the RU-486 is one helix 12 in in the in of GR the and the agonist structures to antagonist from agonist one is of the of helix 12 that in the structure In that of antagonist to the in of helix 12 that of the of RU-486 of helix 12 in the agonist position is seen in the In GR3, helix 11 is at whereas in the helix 11 to of a the structure is out and the is Å and the out the from to a The is with the in other that of the In GR3, the of is to the of the RU-486 is between the two the the between 11 and 12 the other of the dimer and binds in a between the agonist position of helix 12 and the coactivator The of helix 12 in is to that of the position that of helix 12 of the and structures that the helix 12 is to the of the helix 12 position of and the pocket that is has in the of a at the end of helix 11 was is is in the at has the that is from in with of of that and that the of the ligand in and the not GR a of was seen the of between and This of the position of the C terminus of GR in the agonist of GR with dexamethasone and with that seen in is a Å between the helix 12 and the in the part This to that helix 12 the and with C terminus to molecule. This to in the GR with In the of structures with the helix 12 of in the The RU-486 is in the other in The is in that the from the in the of the to and and a the the other the a to and a to the of has to is a that a position in the and antagonist and agonist a different is In the agonist structure is with the ligand and a Å to to position in the In the RU-486 from the ligand and with that in the structure helix 12 a position the ligand-binding pocket, the peptide to in the co-activator a in and the of RU-486 helix 12 from the agonist position in helix 12 is and covers the co-activator the receptor from a is a that has with the of In with GR we the with to in the the the GR to and the was not the was the in the of was to in of in of the was that the of the with a that was to the of GR was This is stretches a In is a of the other pocket between and the GR structure from and structures the type at of with that the a of in the of is in the and a to In the type in and with the of the The in from the structure with the In the type and the a to In the the other out from the pocket and The is and one molecule, of is solved to resolution and not the pocket not in the the and in the The of not to seen a of and of the of a of and 12 in is the coactivator peptide in is The in the structure in and The of not the of GR seen in In was of the the one is a the in the ligand and binds to the agonist structure was solved in in to the structure that was The in the that we two and The dimer in is the to the GR dimer from In GR the of the GR dimer from structure to that GR in the intact of the of the In that of the The the is not in the In the at that the is the and with ligand is in the of the ligand of the agonist structures and that the of the is in the with and at identical the of the in and the the other between different with between in the part of the ligand-binding The in structure between the receptor is seen in in In a to the The position of the of the other in GR-LBD that to the of the between the to a in of GR and and different structures of GR with the antagonist structure helix 12 the part of the coactivator pocket This with is seen in other receptor antagonist In the is 12 together with the end of helix 11 and the out to the agonist and coactivator of a The part of the between 11 and 12 a helix helix the to the position seen in the structure in the agonist structure 12 is the to GR and the of helix The structure is the of the structures together of with other in of different The structures that several to 12 different the is in the 12 is and the coactivator peptide is helix 12 in binds in a position to that of with a of helix 12 and a coactivator peptide in in complex with and crystal The structure was solved in the crystal to Å resolution of and in crystal was not a was that to 2.8 and the was (GR2). In structure the helix 12 was of that to 2.3 Å and with intact C the of The of GR-LBD in and to Å resolution The in The the two crystal forms, and The the has to GR The in the The to a of crystal was the with and The to a of the of coactivator peptide to GR-LBD with The crystal to and the the crystal a and crystal GR3, the one to was solved in a of GR helix in the in the the that was with of and a of to the in was in The the of and to RU-486 in the the the part of the structure was with the in from a and in the The structure was to with and has in and The In the of and in a the agonist structure a to was in with a the structure and the of the structure from the of helix and to with in the was consisting of a dimer and a in the The not the molecule. This with a of the of the of of the from a molecule. with of structure from the the the of a of was out and at of and of a with the This with the that the in the the and with a of the the of the to and the of to and the and and to at the terminus in of the in the The structure is of with and a in crystal is in The GR-LBD structures of of in The structures of and GR to of other in to the receptor and with a of Å and with of Å and 12 not The structures a In GR3, the of helix 12 from the agonist position the ligand-binding pocket in the seen with to the of In the seen of the from of the receptor the RU-486 is one helix 12 in in the in of GR the and the agonist structures to antagonist from agonist one is of the of helix 12 that in the structure In that of antagonist to the in of helix 12 that of the The of RU-486 of helix 12 in the agonist position is seen in the In GR3, helix 11 is at whereas in the helix 11 to of a the structure is out and the is Å and the out the from to a The is with the in other that of the In GR3, the of is to the of the RU-486 is between the two the the between 11 and 12 the other of the dimer and binds in a between the agonist position of helix 12 and the coactivator The of helix 12 in is to that of the position that of helix 12 of the and structures that the helix 12 is to the of the helix 12 position of and the pocket that is has in the of a at the end of helix 11 was is is in the at has the that is from in with of of that and that the of the ligand in and the not GR a of was seen the of between and This of the position of the C terminus of GR in the agonist of GR with dexamethasone and with that seen in is a Å between the helix 12 and the in the part This to that helix 12 the and with C terminus to molecule. This to in the GR with In the of structures with the helix 12 of in the The RU-486 is in the other in The is in that the from the in the of the to and and a the the other the a to and a to the of has to is a that a position in the and antagonist and agonist a different is In the agonist structure is with the ligand and a Å to to position in the In the RU-486 from the ligand and with that in the structure helix 12 a position the ligand-binding pocket, the peptide to in the co-activator a in and the of RU-486 helix 12 from the agonist position in helix 12 is and covers the co-activator the receptor from a is a that has with the of In with GR we the with to in the the the GR to and the was not the was the in the of was to in of in of the was that the of the with a that was to the of GR was This is stretches a In is a of the other pocket between and the GR structure from and structures the type at of with that the a of in the of is in the and a to In the type in and with the of The and in the The of not to seen a of and The agonist structure was solved in in to the structure that was The in the that we two and The dimer in is the to the GR dimer from In GR the of the GR dimer from structure to that GR in the intact of the of the In that of the The the is not in the In the at that the is the and with The ligand is in the of the ligand of the agonist structures and that the of the is in the with and at identical the of the in and the the other between different with between in the part of the ligand-binding The in structure between the receptor is seen in in In a to the The position of the of the other in GR-LBD that to the of the between the to a in different structures of GR with the antagonist structure helix 12 the part of the coactivator pocket This with is seen in other receptor antagonist In the is 12 together with the end of helix 11 and the out to the agonist and coactivator of a The part of the between 11 and 12 a helix helix the to the position seen in the structure in the agonist structure 12 is the to GR and the of helix The structure is the of the structures together of with other in of different The structures that several to The and of at the and the the out with the