Crystal Structure of a Ternary Complex of DnrK, a Methyltransferase in Daunorubicin Biosynthesis, with Bound Products

One of the final steps in the biosynthesis of the widely used anti-tumor drug daunorubicin in Streptomyces peucetius is the methylation of the 4-hydroxyl group of the tetracyclic ring system. This reaction is catalyzed by the S-adenosyl-l-methionine-dependent carminomycin 4-O-methyltransferase DnrK. The crystal structure of the ternary complex of this enzyme with the bound products S-adenosyl-l-homocysteine and 4-methoxy-ϵ-rhodomycin T has been determined to a 2.35-Å resolution. DnrK is a homodimer, and the subunit displays the typical fold of small molecule O-methyltransferases. The structure provides insights into the recognition of the anthracycline substrate and also suggests conformational changes of the of the The and of the bound with of a DnrK enzyme in is to and This the of DnrK with of has been to to One of the final steps in the biosynthesis of the widely used anti-tumor drug daunorubicin in Streptomyces peucetius is the methylation of the 4-hydroxyl group of the tetracyclic ring system. This reaction is catalyzed by the S-adenosyl-l-methionine-dependent carminomycin 4-O-methyltransferase DnrK. The crystal structure of the ternary complex of this enzyme with the bound products S-adenosyl-l-homocysteine and 4-methoxy-ϵ-rhodomycin T has been determined to a 2.35-Å resolution. DnrK is a homodimer, and the subunit displays the typical fold of small molecule O-methyltransferases. The structure provides insights into the recognition of the anthracycline substrate and also suggests conformational changes of the of the The and of the bound with of a DnrK enzyme in is to and This the of DnrK with of has been to to and and widely in the of a of and of a is by in the of the and the of the of with the of the catalyzed by a with of the steps to the of a in the of This is a of in the of by the of of is and the the of daunorubicin biosynthesis in Streptomyces peucetius been The of this anthracycline 4-O-methyltransferase in daunorubicin the methylation of carminomycin the group The of DnrK of and the the is the of used 4-methoxy-ϵ-rhodomycin used 4-methoxy-ϵ-rhodomycin in the biosynthesis of in Streptomyces been to the is is in daunorubicin biosynthesis in Streptomyces and displays a to DnrK. enzyme with is is in biosynthesis in Streptomyces The structure of this determined by the typical fold and a is a and to of this group of has a substrate and in to T the crystal structure of a ternary complex of DnrK with the bound products and 4-methoxy-ϵ-rhodomycin T to a 2.35-Å resolution. The structure provides insights into the recognition of the anthracycline substrate by and also conformational changes of the of the of a by in in This suggests DnrK enzyme in is to and This is the of has been to peucetius by the and the in the the and and to the and DnrK and a to the and with to the DnrK by a in and a in The of of The and the of the in the of the of DnrK and the of the the of the enzyme is the the by the with the and the of The used The products and the of a a of used in a with a a a of with a and daunorubicin by and by T by Streptomyces with the in the by by in The and by of in a reaction and The anthracycline products by and by by and in a reaction and the reaction and the anthracycline products by to the The products by and of the ternary complex of DnrK the of a and in with of used and One crystal in group with of a and and to a of The the to a of The to the of in the crystal of the DnrK ternary complex a in to resolution. The of group with the a and and a of and with the and The of the used group the of the by the and with the of the the in of and structure of in in in in a and structure of the DnrK ternary complex in the crystal by the The of a ternary complex of the and and in The of used a and a and used to the of the in the a with of and of by the of the of The of by and to the to to of the the the fold of the structure by by of by of the the The of and of used to the crystal by the a with a of and of The bound been the and the of the by in the and of this crystal the and the final has of and of of the with and the of the in The and structure been in the with the and and and structure with and the in The and of by DnrK in anthracycline and The carminomycin methylation with and the methylation a with The of this the by the products DnrK with a of the in the of of DnrK and the methylation by the enzyme of the of the by the of the and the with the substrate and the crystal and and to and 2.35-Å The of of the determined with The final is a of in and in and The the and of the The the and the bound is The and with of This is to in the of the of the the bound and in the of DnrK. The is The final of the complex is of a the complex the in and in The the of the group of and the to the 4-hydroxyl of group has The is DnrK in group in and in and in the of the complex in group the the and the bound is a the and in the is the in the of the is by The in group a of the of of the determined in the with The and with of the and the in crystal the is in the the bound This is also by the the bound the the the the of the DnrK ternary complex determined in the crystal of and of the subunit of DnrK is by with a structure a and the fold a by The the the with the The is the and and of in the The fold of DnrK of the to DnrK. with of The in the structure the is in is and of the of the ternary complex of is is and the bound in and structure in the is in the of is by The in and crystal DnrK is a of crystal the with in the crystal in group in The with molecule is of crystal a group the to the and of a The small of this suggests the with in the and of and and the enzyme a of and in with the DnrK the the of the is by The is in the the of the of the and the is bound to DnrK in a in small molecule The with the enzyme and a The ring with the of and and a to the The is to DnrK of the and to the of and with the group the group of and the also a to the of of DnrK and with the of is by in the of the group of to the of to in in is in DnrK. This is by in to in and in of the in the ternary the of the bound with of the The and in in in used with of the of the and of the crystal of the DnrK the of the with the bound to in and bound to in The is in and in The the of to the and of the The the in a the the and to the in crystal a bound anthracycline of the substrate in the into this the to the 4-hydroxyl of the and the of the group of The ternary complex in the a a of the to the enzyme is by and and a the to The of a to the of and with the of The of a molecule with of the to the the of the bound to the One is the a molecule to the of and the is the and the of of of the with the is a of the the bound and the in of the in group the has a is to in this with this the the in the DnrK in this crystal of the of the This is to One a small in the of the with to the of the The is a in the of a in the to the in the crystal and in of the crystal the the of the is and the is to the and a the fold the of the is a and the is in this subunit to with with the of the bound the is bound to the enzyme and is to a conformational changes to substrate and The in the in the crystal conformational changes and to and a of a of in Streptomyces of to DnrK been The to of DnrK is the to is in the biosynthesis of daunorubicin in Streptomyces Streptomyces Streptomyces and a Streptomyces is is group is with the been in the of DnrK and is in the of the 4-hydroxyl group of the substrate of Streptomyces to DnrK. the Streptomyces and and with and the substrate and in been of with a bound in also with a of and a of small to of the fold of the structure a in substrate recognition also the fold and to of DnrK with of and of small molecule and the of DnrK to a of this of a of in the of the ring and the DnrK is of of the ring and is with to the of the the of the with by DnrK. The structure of the DnrK ternary complex provides this of the enzyme to with of the the and and fold the and the the is this and is also to the into the in in anthracycline is a the with in DnrK. The the of the to the and also the substrate to and of and a the group in the enzyme with this The of DnrK by the of the substrate has a to the this this to of a has been DnrK and DnrK also with a group to in the substrate the structure of the ternary complex of the and the of the the of and the of in is the a reaction The structure of DnrK suggests a of group with this of The of One of the of the enzyme to to the of the in group The a by a to the of the of the substrate and the group of the bound products in the of DnrK the of the in the DnrK ternary a of the reaction catalyzed by DnrK is the of the of the 4-hydroxyl group of to the This in in group of the group by the of the group and to has been in the of the and and the and DnrK is in the of the 4-hydroxyl group a of this by in a with suggests is the group of the group also to the The of the group of and the group of the in this is a to by DnrK. This the of this enzyme to of the and a in the reaction DnrK is enzyme and a of DnrK is is in to the and also a and to the and and widely in the of a of and of a is by in the of the and the of the of with the of the catalyzed by a with of the steps to the of a in the of This is a of in the of by the of of is and the The the of daunorubicin biosynthesis in Streptomyces peucetius been The of this anthracycline 4-O-methyltransferase in daunorubicin the methylation of carminomycin the group The of DnrK of and the the is the of used 4-methoxy-ϵ-rhodomycin used 4-methoxy-ϵ-rhodomycin in the biosynthesis of in Streptomyces been to the is is in daunorubicin biosynthesis in Streptomyces and displays a to DnrK. enzyme with is is in biosynthesis in Streptomyces The structure of this determined by the typical fold and a is a and to of this group of DnrK has a substrate and in to T the crystal structure of a ternary complex of DnrK with the bound products and 4-methoxy-ϵ-rhodomycin T to a 2.35-Å resolution. The structure provides insights into the recognition of the anthracycline substrate by and also conformational changes of the of the of a by in in This suggests DnrK enzyme in is to and This is the of has been to peucetius by the and the in the the and and to the and DnrK and a to the and with to the DnrK by a in and a in The of of The and the of the in the of the of DnrK and the of the the of the enzyme is the the by the with the and the of The used The products and the of a a of used in a with a a a of with a and daunorubicin by and by T by Streptomyces with the in the by by in The and by of in a reaction and The anthracycline products by and by by and in a reaction and the reaction and the anthracycline products by to the The products by and of the ternary complex of DnrK the of a and in with of used and One crystal in group with of a and and to a of The the to a of The to the of in the crystal of the DnrK ternary complex a in to resolution. The of group with the a and and a of and with the and The of the used group the of the by the and with the of the the in of and structure of in in in in a and structure of the DnrK ternary complex in the crystal by the The of a ternary complex of the and and in The of used a and a and used to the of the in the a with of and of by the of the of The of by and to the to to of the the the fold of the structure by by of by of the the The of and of used to the crystal by the a with a of and of The bound been the and the of the by in the and of this crystal the and the final has of and of of the with and the of the in The and structure been in the with the and and and structure with and the in The and peucetius by the and the in the the and and to the and DnrK and a to the and with to the DnrK by a in and a in The of of The and the of the in the of the of DnrK and the of the the of the enzyme is the the by the with the and the of The used The products and the of a a of used in a with a a a of with a and daunorubicin by and by T by Streptomyces with the in the by by in The and by of in a reaction and The anthracycline products by and by by and in a reaction and the reaction and the anthracycline products by to the The products by and of the ternary complex of DnrK the of a and in with of used and One crystal in group with of a and and to a of The the to a of The to the of in the crystal of the DnrK ternary complex a in to resolution. The of group with the a and and a of and with the and The of the used group the of the by the and with the of the the in and structure of the DnrK ternary complex in the crystal by the The of a ternary complex of the and and in The of used a and a and used to the of the in the a with of and of The by the of the of The of by and to the to to of the the the fold of the structure by by of by of the the The of and of This used to the crystal by the a with a of and of The bound been the and the of the by in the and of this crystal the and the final has of and of The of the with and the of the in The and structure been in the with the and and and structure with and the in The and of by DnrK in anthracycline and The carminomycin methylation with and the methylation a with The of this the by the products DnrK with a of the in the of of DnrK and the methylation by the enzyme of the of the by the of the and the with the substrate and the crystal and and to and 2.35-Å The of of the determined with The final is a of in and in and The the and of the The the and the bound is The and with of This is to in the of the DnrK in group in and in and in the of the complex in group the the and the bound is a the and in the is the in the of the is by The in group a of the of of the determined in the with The and with of the and the in crystal the is in the the bound This is also by the the bound the the the the of the DnrK ternary complex determined in the crystal of and of the subunit of DnrK is by with a structure a and the fold a by The the the with the The is the and and of in the The fold of DnrK of the to DnrK. with of The in the structure the is in is and of the of the ternary complex of is is and the bound in and structure in the is in the of is by The in and crystal DnrK is a of crystal the with in the crystal in group in The with molecule is of crystal a group the to the and of a The small of this suggests the with in the and of and and the enzyme a of and in with the DnrK the the of the is by The is in the the of the of the and the is bound to DnrK in a in small molecule The with the enzyme and a The ring with the of and and a to the The is to DnrK of the and to the of and with the group the group of and the also a to the of of DnrK and with the of is by in the of the group of to the of to in in is in DnrK. This is by in to in and in of the in the ternary the of the bound with of the The and in in in used with of the of the and of the crystal of the DnrK the of the with the bound to in and bound to in The is in and in The the of to the and of the The the in a the the and to the in crystal a bound anthracycline of the substrate in the into this the to the 4-hydroxyl of the and the of the group of The ternary complex in the a a of the to the enzyme is by and and a the to The of a to the of and with the of The of a molecule with of the to the the of the bound to the One is the a molecule to the of and the is the and the of of of the with the is a of the the bound and the in of the in group the has a is to in this with this the the in the DnrK in this crystal of the of the This is to One a small in the of the with to the of the The is a in the of a in the to the in the crystal and in of the crystal the the of the is and the is to the and a the fold the of the is a and the is in this subunit to with with the of the bound the is bound to the enzyme and is to a conformational changes to substrate and The in the in the crystal conformational changes and to and a of a of in Streptomyces of to DnrK been The to of DnrK is the to is in the biosynthesis of daunorubicin in Streptomyces Streptomyces Streptomyces and a Streptomyces is is group is with the been in the of DnrK and is in the of the 4-hydroxyl group of the substrate of Streptomyces to DnrK. the Streptomyces and and with and the substrate and in been of with a bound in also with a of and a of small to of the fold of the structure a in substrate recognition also the fold and to of DnrK with of and of small molecule and the of DnrK to a of this of of by DnrK in anthracycline and The carminomycin methylation with and the methylation a with The of this the by the products DnrK with a of the in the of of DnrK and the methylation by the enzyme of the of the by the of the and the with the substrate and the crystal and and to and 2.35-Å The of of the determined with The final is a of in and in and The the and of the The the and the bound is The and with of This is to in the of the The DnrK in group in and in and in the of the complex in group the the and the bound is a the and in the is the in the of the is by The in group a of the of The of the determined in the with The and with of the and the in crystal the is in the the bound This is also by the the bound the the the the of the DnrK ternary complex determined in the crystal of and of the subunit of DnrK is by with a structure a and the fold a by The the the with the The is the and and of in the The fold of DnrK of the to DnrK. with of The in the structure the is in is and The is in the of is by The in and crystal DnrK is a of crystal the with in the crystal in group in The with molecule is of crystal a group the to the and of a The small of this suggests the with in the and of and and the enzyme a of and in with the DnrK the the of the is by The is in the the of the of the and the is bound to DnrK in a in small molecule The with the enzyme and a The ring with the of and and a to the The is to DnrK of the and to the of and with the group the group of and the also a to the of of DnrK and with the of is by in the of the group of to the of to in in is in DnrK. This is by in to in a the the and to the in crystal a bound anthracycline of the substrate in the into this the to the 4-hydroxyl of the and the of the group of The ternary complex in the a a The of the to the enzyme is by and and a the to The of a to the of and with the of The of a molecule with of the to the the of the bound to the One is the a molecule to the of and the is the and the of The of of the with the is a of the the bound and the in of the in group the has a is to in this with this the the in the DnrK in this crystal of the of the This is to One a small in the of the with to the of the The is a in the of a in the to the in the crystal and in of the crystal the the of the is and the is to the and a the fold the of the is a and the is in this subunit to with with the of the bound the is bound to the enzyme and is to a conformational changes to substrate and The in the in the crystal conformational changes and to and DnrK a of a of in Streptomyces of to DnrK been The to of DnrK is the to is in the biosynthesis of daunorubicin in Streptomyces Streptomyces Streptomyces and a Streptomyces is is group is with the been in the of DnrK and is in the of the 4-hydroxyl group of the substrate been of with a bound in also with a of and a of small to of the fold of the structure a in substrate recognition also the fold and to of DnrK with of and of small molecule and the of DnrK to a of this of a of in the of the ring and the DnrK is of of the ring and is with to the of the the of the with by DnrK. The structure of the DnrK ternary complex provides this of the enzyme to with of the the and and fold the and the the is this and is also to the into the in in anthracycline is a the with of and a the group in the enzyme with this The of DnrK by the of the substrate has a to the this this to of a has been DnrK and DnrK also with a group to in the substrate the structure of the ternary complex of the and the of the the of and the of in is the a reaction The structure of DnrK suggests a of group with this of The of One of the of the enzyme to to the of the in group The a by a to the of the of the substrate and the group of the bound products in the of DnrK the of the in the DnrK ternary a of the reaction catalyzed by DnrK is the of the of the 4-hydroxyl group of to the This in in group of the group by the of the group and to has been in the of the and and the and DnrK is in the of the 4-hydroxyl group a of this by in a with suggests is the group of the group also to the The of the group of and the group of the in this is a to by DnrK. This the of this enzyme to of the and a in the reaction DnrK is enzyme and a of DnrK is is in to the and also a and to the a of in the of the ring and the DnrK is of of the ring and is with to the of the the of the with by DnrK. The structure of the DnrK ternary complex provides this of the enzyme to with of the the and and fold the and the the is this and is also to the into the in in anthracycline is a the with The of and a the group in the enzyme with this The of DnrK by the of the substrate has a to the this this to of a has been DnrK and DnrK also with a group to in the substrate the structure of the ternary complex of the and the of the the of and the of in is the a reaction The structure of DnrK suggests a of group with this of The of One of the of the enzyme to to the of the in group The a by a to the of the of the substrate and the group the reaction catalyzed by DnrK is the of the of the 4-hydroxyl group of to the This in in group of the group by the of the group and to has been in the of the and and the and DnrK is in the of the 4-hydroxyl group a of this by in a with suggests is the group of the group also to the The of the group of and the group of the in this is a to by DnrK. This the of this enzyme to of the and a in the reaction DnrK is enzyme and a of DnrK is is in to the and also a and to the to and the and