Mutational, Structural, and Kinetic Studies of the ATP-binding Site of Methanobacterium thermoautotrophicum Nicotinamide Mononucleotide Adenylyltransferase

Several residues lining the ATP-binding site of Methanobacterium thermoautotrophicum nicotinamide mononucleotide adenylyltransferase (NMNATase) were mutated in an effort to better characterize their roles in substrate binding and catalysis. Residues selected were Arg-11 and Arg-136, both of which had previously been implicated as substrate binding residues, as well as His-16 and His-19, part of the HXGH active site motif and postulated to be of importance in catalysis. Kinetic studies revealed both Arg-11 and to the binding of the were the of the of and and of and the residues were to of the and were the as of the of the had of the revealed and their active binding of were the binding of in His-16 and and of and of binding in the of and Several residues lining the ATP-binding site of Methanobacterium thermoautotrophicum nicotinamide mononucleotide adenylyltransferase (NMNATase) were mutated in an effort to better characterize their roles in substrate binding and catalysis. Residues selected were Arg-11 and Arg-136, both of which had previously been implicated as substrate binding residues, as well as His-16 and His-19, part of the HXGH active site motif and postulated to be of importance in catalysis. Kinetic studies revealed both Arg-11 and to the binding of the were the of the of and and of and the residues were to of the and were the as of the of the had of the revealed and their active binding of were the binding of in His-16 and and of and of binding in the of and Methanobacterium thermoautotrophicum nicotinamide mononucleotide adenylyltransferase nicotinamide mononucleotide nicotinamide nicotinamide nicotinamide mononucleotide nicotinamide nicotinamide which the of both and and nicotinamide mononucleotide mononucleotide and and Methanobacterium and in both and as of in and which the mononucleotide adenylyltransferase and nicotinamide mononucleotide the thermoautotrophicum of mononucleotide to both an to the of the substrate which the the to to the of to both been in the of the in and been of thermoautotrophicum and were in the the substrate were the of and and the revealed and of and mononucleotide adenylyltransferase of the were to the and the in the of and the their motif their active and the of of to mononucleotide importance of the motif be in the to the of of and the active site motif been and of of the of been in the of their of the of the of the motif to the of of and to the of the of and the of and in the of to the and to the studies revealed residues the the of the the postulated in the of and and of the active site of the of the of binding in which the motif the and the and of as had been the in the binding in the and to characterize the roles of active site residues to be in substrate binding of and and were as previously which of of of were in and and as of the of the of the to and in the of the were in and were in as Kinetic in of and of the of the substrate of to to and of and of to were and of the substrate to of the substrate were and in the as well as the and the and of and were of of the were of and of the were and were and the of the of the were the to the of the of to the and were the and were and in the of in an of in and and in and and and were in the of the of in in the of thermoautotrophicum and as residues were to the roles of active site and residues in binding and of thermoautotrophicum selected residues, His-19, and in Kinetic and were were the of and were the of of as well as of the of and in the and be to the of as and of the residues implicated in the binding of the of were mutated to of in the in the to Arg-11 of thermoautotrophicum and both Arg-11 and to the of binding and thermoautotrophicum Arg-11 the to to in and in in of and and the of an in both to of in both and of Arg-11 and in the of as an the of the of both were to of and and to of the of the to in the of the to the binding of as in the of Arg-11 in the the of of the and the of the of an to of and be in the of the the the of the of in which Arg-11 and in binding the of the part of of the substrate the the to the in binding and and the Arg-11 and were the binding to in both the and the of in the to to the of the of to in of in both the of residues to the binding of the were both binding and of the roles of the residues the active site motif been well of the the motif and of of the residues were of and and the in and in binding the the residues of the motif in and in in in in and to the thermoautotrophicum to the of of in thermoautotrophicum catalysis. in the of the the the the thermoautotrophicum be the to the the the previously the of of to and of of the the the to the be in be the of the thermoautotrophicum the an and and of and of to and their were and were of and of and were to were the of as and and to and of the residues in the of the and the residues residues in the of the in the the as in of residues to of the and to of of to their in of and of in the of the in in the of the of and of in the active as had been of had the to the of and to to to in the active to the well and the in the in the nicotinamide of the of and and the of and the of and and had the site as the of of the binding of the as the as the of the the of the an the and of the of the the of be in the of thermoautotrophicum of the the and to the of in the binding as the as of the of and the to to of the in the of of the the the the the of the the of Arg-11 to of the in both the and binding as both binding in Arg-11 of Arg-136, of the be the in the site to the their to be in the and of had of and the which the of in the ATP-binding site be of the of the to the in the to of to of of the in of the the in the the of and the and the the in the residues in and the and the the in the residues in and to of to of the residues in the of the and the residues residues in the of the of of the of in the residues and of residues of an residues and of the residues an in an the and in the the residues and an were to the of the to of of to their in of the of the revealed of and of the of in the active site of in of to the nicotinamide the part of and to the of Arg-11 and in the of the the of the and the an of of the the of and the the of the of in the of an active site to be the of the of of and of the active site of the and the in of and and roles in binding as well as the and effort to better the of the as well as of in of residues the binding of the were and as well as of the to to and to were in to the the and of the HXGH motif the in of in binding and been as well to of as well as studies of the and postulated the motif binding and the of the residues in an of the active site of to the motif to the and of the and as in of the residues in the motif the of the in the the to the the of binding the and in to of the of the substrate been implicated in catalysis. in binding to of of residues lining the to of the in the residues, Arg-11 and Arg-136, in binding the of His-16 in binding the of in to an in in as in the of the the of the of the and the to the of the residues in the motif the and the the of the the the of the residues of the motif the and of in the the and His-16 the in to in the in which to the in the of thermoautotrophicum an of the which to the of the substrate and of the and of the of the to binding of in the as in the the to to in to the of of the residues the HXGH motif active site in the of Methanobacterium thermoautotrophicum nicotinamide mononucleotide adenylyltransferase nicotinamide mononucleotide nicotinamide nicotinamide nicotinamide mononucleotide nicotinamide nicotinamide which the of both and and nicotinamide mononucleotide mononucleotide and and Methanobacterium and in both and as of in and which the mononucleotide adenylyltransferase and nicotinamide mononucleotide the thermoautotrophicum of mononucleotide to both an to the of the substrate which the the to to the of to both been in the of the in and been of thermoautotrophicum and were in the the substrate were the of and and the revealed and of and mononucleotide adenylyltransferase of the were to the and the in the of and the their motif their active and the of of to mononucleotide importance of the motif be in the to the of of and the active site motif been and of of the of been in the of their of the of the of the motif to the of of and to the of the of and the of and in the of to the and to the studies revealed residues the the of the the postulated in the of and and of the active site of the of the of binding in which the motif the and the and of as had been the in the binding in the and to characterize the roles of active site residues to be in substrate binding of and and were as previously which of of of were in and and as of the of the of the to and in the of the were in and were in as Kinetic in of and of the of the substrate of to to and of and of to were and of the substrate to of the substrate were and in the as well as the and the and of and were of of the were of and of the were and were and the of the of the were the to the of the of to the and were the and were and in the of in an of in and and in and and and were in the of the of in in and and were as previously which of of of were in and and as of the of the of the to and in the of the were in and were in as Kinetic in of and of the of the substrate of to to and of and of to were and of the substrate to Kinetic of the substrate were and in the as well as the and the and of and were of of the were of and of the were and were and the of the of the were the to the of the of to the and were the and were and in the of in an of in and and in and and and were in the of the the of thermoautotrophicum and as residues were to the roles of active site and residues in binding and of thermoautotrophicum selected residues, His-19, and in Kinetic and were were the of and were the of of as well as of the of and in the and be to the of as and of the residues implicated in the binding of the of were mutated to of in the in the to Arg-11 of thermoautotrophicum and both Arg-11 and to the of binding and thermoautotrophicum Arg-11 the to to in and in in of and and the of an in both to of in both and of Arg-11 and in the of as an the of the of both were to of and and to of the of the to in the of the to the binding of as in the of Arg-11 in the the of of the and the of the of an to of and be in the of the the the of the of in which Arg-11 and in binding the of the part of of the substrate the the to the in binding and and the Arg-11 and were the binding to in both the and the of in the to to the of the of to in of in both the of residues to the binding of the were both binding and of the roles of the residues the active site motif been well of the the motif and of of the residues were of and and the in and in binding the the residues of the motif in and in in in in and to the thermoautotrophicum to the of of in thermoautotrophicum catalysis. in the of the the the the thermoautotrophicum be the to the the the previously the of of to and of of the the the to the be in be the of the thermoautotrophicum the an and and of and of to and their were and were of and of and were to were the of as and and to and of the residues in the of the and the residues residues in the of the in the the as in of residues to of the and to of of to their in of and of in the of the in in the of the of and of in the active as had been of had the to the of and to to to in the active to the well and the in the in the nicotinamide of the of and and the of and the of and and had the site as the of of the binding of the as the as the of the the of the an the and of the of the the of be in the of thermoautotrophicum of the the and to the of in the binding as the as of the of and the to to of the in the of of the the the the the of the the of Arg-11 to of the in both the and binding as both binding in Arg-11 of Arg-136, of the be the in the site to the their to be in the and of had of and the which the of in the ATP-binding site be of the of the to the in the to of to of of the in of the the in the the of and the and the the in the residues in and to of to of the residues in the of the and the residues residues in the of the of of the of in the residues and of residues of an residues and of the residues an in an the and in the the residues and an were to the of the to of of to their in of the of the revealed of and of the of in the active site of in of to the nicotinamide the part of and to the of Arg-11 and in the of the the of the and the an of of the the of and the the of the of in the of an active site to be the of the of of and of the active site of the and the in of and and roles in binding as well as the and effort to better the of the as well as of in of residues the binding of the were and as well as of the to to and to were in to the the and of the HXGH motif the in of in binding and been as well to of as well as studies of the and postulated the motif binding and the of the residues in an of the active site of to the motif to the and of the and as in of the residues in the motif the of the in the the to the the of binding the and in to of the of the substrate been implicated in catalysis. in binding to of of residues lining the to of the in the residues, Arg-11 and Arg-136, in binding the of His-16 in binding the of in to an in in as in the of the the of the of the and the to the of the residues in the motif the and the the of the the the of the residues of the motif the and of in the the and His-16 the in to in the in which to the in the of thermoautotrophicum an of the which to the of the substrate and of the and of the of the to binding of in the as in the the to to in to the of of the residues the HXGH motif active site in the of the of thermoautotrophicum and as residues were to the roles of active site and residues in binding and of thermoautotrophicum selected residues, His-19, and in the of thermoautotrophicum and as residues were to the roles of active site and residues in binding and of thermoautotrophicum selected residues, His-19, and in Kinetic and were were the of and were the of of as well as of the of and in and were were the of and were the of of as well as of the the and be to the of as and of the residues implicated in the binding of the of were mutated to of in the in the to Arg-11 of thermoautotrophicum and both Arg-11 and to the of binding and thermoautotrophicum Arg-11 the to to in and in in of and and the of an in both to of in both and of Arg-11 and in the of as an the of the of both were to of and and to of the of the to in the of the to the binding of as in the of Arg-11 in the the of of the and the of the of an to of and be in the of the the the of the of in which Arg-11 and in binding the of the part of of the substrate the the to the in binding and and the Arg-11 and were the binding to in both the and the of in the to to the of the of to in of in both the of residues to the binding of the were both binding and of the roles of the residues the active site motif been well of the the motif and of of the residues were of and and the in and in binding the the residues of the motif in and in in in in and to the thermoautotrophicum to the of of in thermoautotrophicum catalysis. in the of the the the the thermoautotrophicum be the to the the the previously the of of to and of of the the the to the be in be the of the thermoautotrophicum the an and and be to the of as and of the residues implicated in the binding of the of were mutated to of in the in the to Arg-11 of thermoautotrophicum and both Arg-11 and to the of binding and thermoautotrophicum Arg-11 the to to in and in in of and and the of an in both to of in both and of Arg-11 and in the of as an the of the of both were to of and and to of the of the to in the of the to the binding of as in the of Arg-11 in the the of of the and the of the of an to of and be in the of the the the of the of in which Arg-11 and in binding the of the part of of the substrate the the to the in binding and and the Arg-11 and were the binding to in both the and the of in the to to the of the of to in of in both the of residues to the binding of the were both binding catalysis. and of the roles of the residues the active site motif been well of the the motif and of of the residues were of and and the in and in binding the the residues of the motif in and in in in in and to the thermoautotrophicum to the of of in thermoautotrophicum catalysis. in the of the the the the thermoautotrophicum be the to the the the previously the of of to and of of the the the to the be in be the of the thermoautotrophicum the an and and of and of to and their were and were of and of and were to were the of as and and to and of the residues in the of the and the residues residues in the of the in the the as in of residues to of the and to of of to their in of and of in the of the in in the of the of and of in the active as had been of had the to the of and to to to in the active to the well and the in the in the nicotinamide of the of and and the of and the of and and had the site as the of of the binding of the as the as the of the the of the an the and of the of the the of be in the of thermoautotrophicum of the the and to the of in the binding as the as of the of and the to to of the in the of of the the the the the of the the of Arg-11 to of the in both the and binding as both binding in Arg-11 of Arg-136, of the be the in the site to the their to be in the and of had of and the which the of in the ATP-binding site be of the of the to the in the to of to of of the in of the the in the the of and to of to of the residues in the of the and the residues residues in the of the of of the of in the residues and of residues of an residues and of the residues an in an the and in the the residues and an were to the of the to of of to their in of the of the revealed of and of the of in the active site of in of to the nicotinamide the part of and to the of Arg-11 and in the of the the of the and the an of of the the of and the the of the of in the of an active site to be the of the of of and of the active site of the and the in of and and roles in binding as well as the and effort to better the of the as well as of in of residues the binding of the were and as well as of the to to and to were in to the the and of the HXGH motif the in of in binding and been as well to of as well as studies of the and postulated the motif binding and the of the residues in an of the active site of to the motif to the and of the and as in of the residues in the motif the of the in the the to the the of binding the and in to of the of the substrate been implicated in catalysis. in binding to of of residues lining the to of the in the residues, Arg-11 and Arg-136, in binding the of His-16 in binding the of in to an in in as in the of the the of the of the and the to the of the residues in the motif the and the the of the the the of the residues of the motif the and of in the the and His-16 the in to in the in which to the in the of thermoautotrophicum an of the which to the of the substrate and of the and of the of the to binding of in the as in the the to to in to the of of the residues the HXGH motif active site in the of and of and of to and their were and were of and of and were to were the of as and and to and of the residues in the of the and the residues residues in the of the in the the as in of residues to of the and to of of to their in of and of in the of the in in the of the of and of in the active as had been of had the to the of and to to to in the active to the well and the in the in the nicotinamide of the of and and the of and the of and and had the site as the of of the binding of the as the as the of the the of the an the and of the of the the of be in the of thermoautotrophicum of the the and to the of in the binding as the as of the of and the to to of the in the of of the the the the the of the the of Arg-11 to of the in both the and binding as both binding in Arg-11 of Arg-136, of the be the in the site to the their to be in the and of had of and the which the of in the ATP-binding site be of the of the to the in the to of to of of the in of the the in the the of and to of to of the residues in the of the and the residues residues in the of the of of the of in the residues and of residues of an residues and of the residues an in an the and in the the residues and an were to the of the to of of to their in of the of the revealed of and of the of in the active site of in of to the nicotinamide the part of and to the of Arg-11 and in the of the the of the and the an of of the the of and the the of the of in the of an active site to be the of the effort to better the of the as well as of in of residues the binding of the were and as well as of the to to and to were in to the the and of the HXGH motif the in of in binding and been as well to of as well as studies of the and postulated the motif binding and the of the residues in an of the active site of to the motif to the and of the and as in of the residues in the motif the of the in the the to the the of binding the and in to of the of the substrate been implicated in catalysis. in binding to of of residues lining the to of the in the residues, Arg-11 and Arg-136, in binding the of His-16 in binding the of in to an in in as in the of the the of the of the and the to the of the residues in the motif the and the the of the the the of the residues of the motif the and of in the the and His-16 the in to in the in which to the in the of thermoautotrophicum an of the which to the of the substrate and of the and of the of the to binding of in the as in the the to to in to the of of the residues the HXGH motif active site in the of of of the as well as and of the to and of

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.