The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants
<p>Farnesyl pyrophosphate synthase (FPPS) is the major molecular target of nitrogen-containing bisphosphonates (N-BPs), used clinically as bone resorption inhibitors. We investigated the role of threonine 201 (Thr201) and tyrosine 204 (Tyr204) residues in substrate binding, catalysis and inhib...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Elsevier
2015
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| _version_ | 1797064380469215232 |
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| author | Tsoumpra, MK Muniz, JR Barnett, BL Kwaasi, AA Pilka, ES Kavanagh, KL Evdokimov, A Walter, RL Von Delft, F Ebetino, FH Oppermann, U Russell, RGG Dunford, JE |
| author_facet | Tsoumpra, MK Muniz, JR Barnett, BL Kwaasi, AA Pilka, ES Kavanagh, KL Evdokimov, A Walter, RL Von Delft, F Ebetino, FH Oppermann, U Russell, RGG Dunford, JE |
| author_sort | Tsoumpra, MK |
| collection | OXFORD |
| description | <p>Farnesyl pyrophosphate synthase (FPPS) is the major molecular target of nitrogen-containing bisphosphonates (N-BPs), used clinically as bone resorption inhibitors. We investigated the role of threonine 201 (Thr201) and tyrosine 204 (Tyr204) residues in substrate binding, catalysis and inhibition by N-BPs, employing kinetic and crystallographic studies of mutated FPPS proteins.</p>
<p>Mutants of Thr201 illustrated the importance of the methyl group in aiding the formation of the Isopentenyl pyrophosphate (IPP) binding site, while Tyr204 mutations revealed the unknown role of this residue in both catalysis and IPP binding. The interaction between Thr201 and the side chain nitrogen of N-BP was shown to be important for tight binding inhibition by zoledronate (ZOL) and risedronate (RIS), although RIS was also still capable of interacting with the main-chain carbonyl of Lys200. The interaction of RIS with the phenyl ring of Tyr204 proved essential for the maintenance of the isomerized enzyme-inhibitor complex. Studies with conformationally restricted analogues of RIS reaffirmed the importance of Thr201 in the formation of hydrogen bonds with N-BPs.</p>
<p>In conclusion we have identified new features of FPPS inhibition by N-BPs and revealed unknown roles of the active site residues in catalysis and substrate binding.</p> |
| first_indexed | 2024-03-06T21:13:27Z |
| format | Journal article |
| id | oxford-uuid:3ef4e8d0-4bc8-4280-a1d0-2cf9e6a81b1c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:13:27Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:3ef4e8d0-4bc8-4280-a1d0-2cf9e6a81b1c2022-03-26T14:29:00ZThe inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutantsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:3ef4e8d0-4bc8-4280-a1d0-2cf9e6a81b1cEnglishORA DepositElsevier2015Tsoumpra, MKMuniz, JRBarnett, BLKwaasi, AAPilka, ESKavanagh, KLEvdokimov, AWalter, RLVon Delft, FEbetino, FHOppermann, URussell, RGGDunford, JE<p>Farnesyl pyrophosphate synthase (FPPS) is the major molecular target of nitrogen-containing bisphosphonates (N-BPs), used clinically as bone resorption inhibitors. We investigated the role of threonine 201 (Thr201) and tyrosine 204 (Tyr204) residues in substrate binding, catalysis and inhibition by N-BPs, employing kinetic and crystallographic studies of mutated FPPS proteins.</p> <p>Mutants of Thr201 illustrated the importance of the methyl group in aiding the formation of the Isopentenyl pyrophosphate (IPP) binding site, while Tyr204 mutations revealed the unknown role of this residue in both catalysis and IPP binding. The interaction between Thr201 and the side chain nitrogen of N-BP was shown to be important for tight binding inhibition by zoledronate (ZOL) and risedronate (RIS), although RIS was also still capable of interacting with the main-chain carbonyl of Lys200. The interaction of RIS with the phenyl ring of Tyr204 proved essential for the maintenance of the isomerized enzyme-inhibitor complex. Studies with conformationally restricted analogues of RIS reaffirmed the importance of Thr201 in the formation of hydrogen bonds with N-BPs.</p> <p>In conclusion we have identified new features of FPPS inhibition by N-BPs and revealed unknown roles of the active site residues in catalysis and substrate binding.</p> |
| spellingShingle | Tsoumpra, MK Muniz, JR Barnett, BL Kwaasi, AA Pilka, ES Kavanagh, KL Evdokimov, A Walter, RL Von Delft, F Ebetino, FH Oppermann, U Russell, RGG Dunford, JE The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title | The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title_full | The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title_fullStr | The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title_full_unstemmed | The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title_short | The inhibition of human farnesyl pyrophosphate synthase by nitrogen-containing bisphosphonates. Elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
| title_sort | inhibition of human farnesyl pyrophosphate synthase by nitrogen containing bisphosphonates elucidating the role of active site threonine 201 and tyrosine 204 residues using enzyme mutants |
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