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

Overview

Journal Bone

Specialties Endocrinology
Orthopedics

Date 2015 Aug 31

PMID 26318908

Citations 28

Authors

Maria K Tsoumpra

Joao R Muniz

Bobby L Barnett

Aaron A Kwaasi

Ewa S Pilka

Kathryn L Kavanagh

Artem Evdokimov

Richard L Walter

Frank von Delft

Frank H Ebetino

Udo Oppermann

R Graham G Russell

James E Dunford

Affiliations

Soon will be listed here.

Abstract

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. 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. 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.

Citing Articles

Hydroxychloroquine and a low antiresorptive activity bisphosphonate conjugate prevent and reverse ovariectomy-induced bone loss in mice through dual antiresorptive and anabolic effects.

Yao Z, Ayoub A, Srinivasan V, Wu J, Tang C, Duan R Bone Res. 2024; 12(1):52.

PMID: 39231935 PMC: 11375055. DOI: 10.1038/s41413-024-00352-6.

Sclerostin and Wnt Signaling in Idiopathic Juvenile Osteoporosis Using High-Resolution Confocal Microscopy for Three-Dimensional Analyses.

Pereira R, Noche K, Gales B, Chen Z, Salusky I, Albrecht L Children (Basel). 2024; 11(7).

PMID: 39062269 PMC: 11276078. DOI: 10.3390/children11070820.

Thonzonium bromide inhibits progression of malignant pleural mesothelioma through regulation of ERK1/2 and p38 pathways and mitochondrial uncoupling.

DellAnno I, Morani F, Patergnani S, Daga A, Pinton P, Giorgi C Cancer Cell Int. 2024; 24(1):226.

PMID: 38951927 PMC: 11218145. DOI: 10.1186/s12935-024-03400-7.

Antimicrobial and Anesthetic Niosomal Formulations Based on Amino Acid-Derived Surfactants.

Romeo M, Hafidi Z, Muzzalupo R, Pons R, Garcia M, Mazzotta E Molecules. 2024; 29(12).

PMID: 38930908 PMC: 11206639. DOI: 10.3390/molecules29122843.

Hydroxychloroquine and a low activity bisphosphonate conjugate prevent and reverse ovariectomy-induced bone loss in mice through dual antiresorptive and anabolic effects.

Yao Z, Ayoub A, Srinivasan V, Wu J, Tang C, Duan R Res Sq. 2024; .

PMID: 38746138 PMC: 11092802. DOI: 10.21203/rs.3.rs-4237258/v1.

References

Coxon F, Helfrich M, vant Hof R, Sebti S, Ralston S, Hamilton A . Protein geranylgeranylation is required for osteoclast formation, function, and survival: inhibition by bisphosphonates and GGTI-298. J Bone Miner Res. 2000; 15(8):1467-76. DOI: 10.1359/jbmr.2000.15.8.1467. View

Sanz-Rodriguez C, Concepcion J, Pekerar S, Oldfield E, Urbina J . Bisphosphonates as inhibitors of Trypanosoma cruzi hexokinase: kinetic and metabolic studies. J Biol Chem. 2007; 282(17):12377-87. DOI: 10.1074/jbc.M607286200. View

Luckman S, Coxon F, Ebetino F, Russell R, Rogers M . Heterocycle-containing bisphosphonates cause apoptosis and inhibit bone resorption by preventing protein prenylation: evidence from structure-activity relationships in J774 macrophages. J Bone Miner Res. 1998; 13(11):1668-78. DOI: 10.1359/jbmr.1998.13.11.1668. View

Ebetino F, Hogan A, Sun S, Tsoumpra M, Duan X, Triffitt J . The relationship between the chemistry and biological activity of the bisphosphonates. Bone. 2011; 49(1):20-33. DOI: 10.1016/j.bone.2011.03.774. View

Eastell R, Walsh J, Watts N, Siris E . Bisphosphonates for postmenopausal osteoporosis. Bone. 2011; 49(1):82-8. DOI: 10.1016/j.bone.2011.02.011. View

Evans P . Scaling and assessment of data quality. Acta Crystallogr D Biol Crystallogr. 2005; 62(Pt 1):72-82. DOI: 10.1107/S0907444905036693. View

McCoy A, Grosse-Kunstleve R, Adams P, Winn M, Storoni L, Read R . Phaser crystallographic software. J Appl Crystallogr. 2009; 40(Pt 4):658-674. PMC: 2483472. DOI: 10.1107/S0021889807021206. View

Brocklehurst K . A sound basis for pH-dependent kinetic studies on enzymes. Protein Eng. 1994; 7(3):291-9. DOI: 10.1093/protein/7.3.291. View

Artz J, Wernimont A, Dunford J, Schapira M, Dong A, Zhao Y . Molecular characterization of a novel geranylgeranyl pyrophosphate synthase from Plasmodium parasites. J Biol Chem. 2010; 286(5):3315-22. PMC: 3030337. DOI: 10.1074/jbc.M109.027235. View

10.

Reid I, Hosking D . Bisphosphonates in Paget's disease. Bone. 2010; 49(1):89-94. DOI: 10.1016/j.bone.2010.09.002. View

11.

Powell H . The Rossmann Fourier autoindexing algorithm in MOSFLM. Acta Crystallogr D Biol Crystallogr. 1999; 55(Pt 10):1690-5. DOI: 10.1107/s0907444999009506. View

12.

Hosfield D, Zhang Y, Dougan D, Broun A, Tari L, Swanson R . Structural basis for bisphosphonate-mediated inhibition of isoprenoid biosynthesis. J Biol Chem. 2003; 279(10):8526-9. DOI: 10.1074/jbc.C300511200. View

13.

Morgan G, Davies F, Gregory W, Cocks K, Bell S, Szubert A . First-line treatment with zoledronic acid as compared with clodronic acid in multiple myeloma (MRC Myeloma IX): a randomised controlled trial. Lancet. 2010; 376(9757):1989-99. PMC: 3639680. DOI: 10.1016/S0140-6736(10)62051-X. View

14.

Varela I, Pereira S, Ugalde A, Navarro C, Suarez M, Cau P . Combined treatment with statins and aminobisphosphonates extends longevity in a mouse model of human premature aging. Nat Med. 2008; 14(7):767-72. DOI: 10.1038/nm1786. View

15.

Hounslow A, Carran J, Brown R, Rejman D, Blackburn G, Watts D . Determination of the microscopic equilibrium dissociation constants for risedronate and its analogues reveals two distinct roles for the nitrogen atom in nitrogen-containing bisphosphonate drugs. J Med Chem. 2008; 51(14):4170-8. DOI: 10.1021/jm7015792. View

16.

Poulter C, RILLING H . Prenyltransferase: the mechanism of the reaction. Biochemistry. 1976; 15(5):1079-83. DOI: 10.1021/bi00650a019. View

17.

Dunford J, Thompson K, Coxon F, Luckman S, Hahn F, Poulter C . Structure-activity relationships for inhibition of farnesyl diphosphate synthase in vitro and inhibition of bone resorption in vivo by nitrogen-containing bisphosphonates. J Pharmacol Exp Ther. 2001; 296(2):235-42. View

18.

Williams J, Morrison J . The kinetics of reversible tight-binding inhibition. Methods Enzymol. 1979; 63:437-67. DOI: 10.1016/0076-6879(79)63019-7. View

19.

Rondeau J, Bitsch F, Bourgier E, Geiser M, Hemmig R, Kroemer M . Structural basis for the exceptional in vivo efficacy of bisphosphonate drugs. ChemMedChem. 2006; 1(2):267-73. DOI: 10.1002/cmdc.200500059. View

20.

Buhaescu I, Izzedine H . Mevalonate pathway: a review of clinical and therapeutical implications. Clin Biochem. 2007; 40(9-10):575-84. DOI: 10.1016/j.clinbiochem.2007.03.016. View