Abstract
Adenosylmethionine synthetase plays a key role in the biogenesis of the sulfonium compound S-adenosylmethionine, the principal widely used methyl donor in the biological methylations. We report here, for the first time, the characterization of adenosylmethionine synthetase from the hyperthermophilic archaeon Pyrococcus furiosus (PfMAT). The gene PF1866 encoding PfMAT was cloned and expressed, and the recombinant protein was purified to homogeneity. PfMAT shares 51, 63, and 82 % sequence identity with the homologous enzymes from Sulfolobus solfataricus, Methanococcus jannaschii, and Thermococcus kodakarensis, respectively. PfMAT is a homodimer of 90 kDa highly thermophilic with an optimum temperature of 90 °C and is characterized by remarkable thermodynamic stability (Tm, 99 °C), kinetic stability, and resistance to guanidine hydrochloride-induced unfolding. The latter process is reversible as demonstrated by the analysis of the refolding process by activity assays and fluorescence measurements. Limited proteolysis experiments indicated that the proteolytic cleavage site is localized at Lys148 and that the C-terminal peptide is necessary for the integrity of the active site. PfMAT shows kinetic features that make it the most efficient catalyst for S-adenosylmethionine synthesis among the characterized MAT from Bacteria and Archaea. Molecular and structural characterization of PfMAT could be useful to improve MAT enzyme engineering for biotechnological applications.
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Abbreviations
- AdoMet:
-
S-Adenosylmethionine
- MAT:
-
Adenosylmethionine synthetase or methionine adenosyltransferase
- PfMAT:
-
MAT from Pyrococcus furiosus
- SsMAT:
-
MAT from Sulfolobus solfataricus
- MjMAT:
-
MAT from Methanococcus jannaschii
- TkMAT:
-
MAT from Thermococcus kodakarensis
- EcMAT:
-
MAT from Escherichia coli
- BsMAT:
-
MAT from Bacillus subtilis
- GdnCl:
-
Guanidine hydrochloride
- IPTG:
-
Isopropyl-β-d-thiogalactopyranoside
- PVDF:
-
Polyvinylidene fluoride
- CD:
-
Circular dichroism
References
Salvatore, F., Borek, E., Zappia, V., Williams-Ashman, H. G., & Schlenk, F. (1977). The biochemistry of adenosylmethionine. New York: Columbia University Press.
Usdin, E., Borchardt, C. R. T., & Creveling, R. (1982). Biochemistry of s-adenosylmethionine and related compounds. London: MacMillan Press.
Lu, S. C. (2000). S-Adenosylmethionine. International Journal of Biochemistry and Cell Biology, 32, 391–395.
Mato, J. M., Corrales, F. J., Lu, S. C., & Avila, M. A. (2002). S-Adenosylmethionine: a control switch that regulates liver function. FASEB Journal, 16, 15–26.
Fontecave, M., Atta, M., & Mulliez, E. (2004). S-Adenosylmethionine: nothing goes to waste. Trends Biochemical Sciences, 29, 243–249.
Cantoni, G. L. (1953). S-Adenosylmethionine, a new intermediate formed enzimatically from L-metionine and adenosinetriphosphate. Journal of Biological Chemistry, 203, 403–416.
Chiang, P. K., Gordon, R. K., Tal, J., Zeng, G. C., Doctor, B. P., Pardhasaradhi, K., & McCann, P. P. (1996). S-Adenosylmethionine and methylation. FASEB Journal, 10, 471–480.
Lu, S. C. (2009). Regulation of glutathione synthesis. Molecular Aspects of Medicine, 30, 42–59.
Frey, P. A., Hegeman, A. D., & Ruzicka, F. J. (2008). The radical SAM superfamily. Critical Reviews in Biochemistry and Molecular Biology, 43, 63–88.
Sauter, M., Moffatt, B., Saechao, M. C., Hell, R., & Wirtz, M. (2013). Methionine salvage and S-adenosylmethionine: essential links between sulfur, ethylene and polyamine biosynthesis. Biochemistry Journal, 451, 145–154.
Ansorena, E., García-Trevijano, E. R., Martínez-Chantar, M. L., Huang, Z. Z., Chen, L., Mato, J. M., Iraburu, M., Lu, S. C., & Avila, M. A. (2002). S-Adenosylmethionine and methylthioadenosine are antiapoptotic in cultured rat hepatocytes but proapoptotic in human hepatoma cells. Hepatology, 35, 274–280.
Martinez-Lopez, N., Valera-Rey, M., Ariz, U., Embade, N., Vazquez-Chantada, M., Fernandez-Ramos, D., Gomez-Santos, L., Lu, S. C., Mato, J. M., & Martinez-Chantar, M. L. (2008). S-Adenosylmethionine and proliferation: new pathways, new targets. Biochemical Society Transactions, 36, 848–852.
Li, T. W. H., Yang, H., Peng, H., Xia, M., Mato, J. M., & Lu, S. C. (2012). Effects of S-adenosylmethionine and metylthioadenosine on inflammation-induced colon cancer in mice. Carcinogenesis, 33, 427–435.
Bottiglieri, T. (2002). S-Adenosyl-L-methionine (SAMe): from the bench to the bedside-molecular basis of a pleiotrophic molecule. American Journal of Clinical Nutrition, 76, 1151S–1157S.
Papakostas, G. I., Cassiello, C. F., & Iovieno, N. (2012). Folates and S-adenosylmethionine for major depressive disorder. Canadian Journal of Psychiatry. Revue Canadienne de Psychiatrie, 57, 406–413.
Soeken, K. L., Lee, W. L., Bausell, R. B., Agelli, M., & Berman, B. M. (2012). Safety and efficacy of S-adenosylmethionine (SAMe) for osteoarthritis. Journal Farmacia Practice, 51, 425–430.
Anstee, Q. M., & Day, C. P. (2012). S-Adenosylmethionine (SAMe) therapy in liver disease: a review of current evidence and clinical utility. Journal of Hepatology, 57, 1097–1109.
Lu, S. C., & Mato, J. M. (2012). S-Adenosylmethionine in liver health, injury, and cancer. Physiological Reviews, 92, 1515–1542.
Mato, J. M., Martinez-Chantar, M. L., & Lu, S. C. (2013). S-Adenosylmethionine metabolism and liver disease. Annals of Hepatology, 12, 183–189.
Kotb, M., & Geller, A. M. (1993). Methionine adenosyltransferase: structure and function. Pharmacology and Therapeutics, 59, 125–143.
Mato, J. M., Alvarez, L., Ortiz, P., & Pajares, M. A. (1997). S-Adenosylmethionine synthesis: molecular mechanisms and clinical implications. Pharmacology and Therapeutics, 73, 265–280.
Markham, G. D., & Pajares, M. A. (2009). Structure-function relationships in methionine adenosyltransferases. Cellular and Molecular Life Sciences, 66, 636–648.
Pajares, M. A., & Markham, G. D. (2011). Methionine adenosyltransferase (S-adenosyl-methionine synthetase). Advances in Enzymology and Related Areas of Molecular Biology, 78, 449–452.
Markham, G. D., Hafner, E. W., Tabor, C. W., & Tabor, H. (1980). S-Adenosylmethionine synthetase from Escherichia coli. Journal of Biological Chemistry, 255, 9082–9092.
Komoto, J., Yamada, T., Takata, Y., Markham, G. D., & Takusagawa, F. (2004). Crystal structure of the S-adenosylmethionine synthetase ternary complex: a novel catalytic mechanism of S-adenosylmethionine synthesis from ATP and Met. Biochemistry, 43, 1821–1831.
Gonzalez, B., Pajares, M. A., Hermoso, J. A., Guillerm, D., Guillerm, G., & Sanz-Aparicio, J. (2003). Crystal structures of methionine adenosyltransferase complexed with substrates and products reveal the methionine-ATP recognition and give insights into the catalytic mechanism. Journal of Molecular Biology, 331, 407–416.
Alvarez, L., Corrales, F., Martin-Duce, A., & Mato, J. M. (1993). Characterization of a full-length cDNA encoding human liver S-adenosylmethionine synthetase: tissue-specific gene expression and mRNA levels in hepatopathies. Biochemistry Journal, 293, 481–486.
Shafqat, N., Muniz, J. R., Pilka, E. S., Papagrigoriou, E., vonDelft, F., Oppermann, U., & Yue, W. W. (2013). Insight into S-adenosylmethionine biosynthesis from the crystal structures of the human methionine adenosyltransferase catalytic and regulatory subunits. Biochemistry Journal, 452, 27–36.
Reytor, E., Pérez-Miguelsanz, J., Alvarez, L., Pèrez-Sala, D., & Pajares, M. A. (2009). Conformational signals in the C-terminal domain of methionine adenosyltransferase I/III determine its nucleocytoplasmic distribution. FASEB Journal, 23, 3347–3360.
De Rosa, M., De Rosa, S., Gambacorta, A., Cartenì-Farina, M., & Zappia, V. (1978). The biosynthetic pathway of new polyamines in Caldariella acidophila. Biochemistry Journal, 176, 1–7.
Porcelli, M., Cacciapuoti, G., Cartenì-Farina, M., & Gambacorta, A. (1988). S-Adenosylmethionine synthetase in the thermophilic archaebacterium Sulfolobus solfataricus. Purification and characterization of two isoforms. European Journal of Biochemistry, 177, 273–280.
Graham, D. E., Bock, C. L., Schalk-Hihi, C., Lu, Z. J., & Markham, G. D. (2000). Identification of a highly diverged class of S-adenosylmethionine synthetases in the Archaea. Journal of Biological Chemistry, 275, 4055–4059.
Lu, Z. J., & Markham, G. D. (2002). Enzymatic properties of S-adenosylmethionine synthetase from the archaeon Methanococcus jannaschii. Journal of Biological Chemistry, 277, 16624–16631.
Garrido, F., Alfonso, C., Taylor, J. C., Markham, G. D., & Pajares, M. A. (2009). Subunit association as the stabilizing determinant for archaeal methionine adenosyltransferases. Biochimica et Biophysica Acta, 1794, 1082–1090.
Garrido, F., Taylor, J. C., Alfonso, C., Markham, G. D., & Pajares, M. A. (2012). Structural basis for the stability of a thermophilic methionine adenosyltransferase against guanidinium chloride. Amino Acids, 42, 361–373.
Schlesier, J., Siegrist, J., Gerhardt, S., Erb, A., Blaesi, S., Richter, M., Einsle, O., & Andexer, J. N. (2013). Structural and functional characterization of the methionine adenosyltransferase from Thermococcus kodakarensis. BMC Structural Biology, 13, 22–31.
Wang, F., Singh, S., Zhang, J., Huber, T. D., Helmich, K. E., Sunkara, M., Hurley, K. A., Goff, R. D., Bingman, C. A., Morris, A. J., Thorson, J. S., & Phillips, G. N., Jr. (2014). Understanding molecular recognition of promiscuity of thermophilic methionineadenosyltransferase, sMAT from Sulfolobus solfataricus. FEBS Journal. doi:10.1111/febs.12784.
Adams, M. W. W., & Kelly, R. M. (1994). Thermostability and thermoactivity of enzymes from hyperthermophilic Archaea. Bioorganic & Medicinal Chemistry, 2, 659–667.
Vieille, C., & Zeikus, G. J. (2001). Hyperthermophilic enzymes: sources, uses and molecular mechanisms for thermostability. Microbiology and Molecular Biology Reviews, 65, 1–43.
Niehaus, F., Bertoldo, C., Kahler, M., & Antranikian, G. (1999). Extremophiles as a source of novel enzymes for industrial application. Applied Microbiology and Biotechnology, 51, 711–729.
Fiala, G., & Stetter, K. O. (1986). Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100 °C. Archives of Microbiology, 145, 56–61.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.
Cacciapuoti, G., Porcelli, M., Bertoldo, C., De Rosa, M., & Zappia, V. (1994). Purification and characterization of extremely thermophilic and thermostable 5′-methylthioadenosine phosphorylase from the archaeon Sulfolobus solfataricus. Purine nucleoside phosphorylase activity and evidence for intersubunit disulfide bonds. Journal of Biological Chemistry, 269, 24762–24769.
Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Laboratory.
Cacciapuoti, G., Fuccio, F., Petraccone, L., Del Vecchio, P., & Porcelli, M. (2012). Role of disulfide bonds in conformational stability and folding of 5-deoxy-5-methylthioadenosine phosphorylase II from the hyperthermophilic archaeon Sulfolobus solfataricus. Biochimica et Biophysica Acta, 1824, 1136–1143.
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., & Higgins, D. G. (1997). The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876–4882.
Ragone, R., Facchiano, F., Cacciapuoti, G., Porcelli, M., & Colonna, G. (1992). Effect of temperature on the propylamine transferase from Sulfolobus solfataricus, an extreme thermophilic archaebacterium. 2. Denaturation and structural stability. European Journal of Biochemistry, 204, 483–490.
Faraone Mennella, M. R., Gambacorta, A., Nicolaus, B., & Farina, B. (1998). Purification and biochemical characterization of a poly(ADP-ribose) polymearse-like enzyme from the thermophilic archaeon Sulfolobus solfataricus. Biochemistry Journal, 335, 441–447.
Kamarthapu, V., Rao, K. V., Srinivas, B. S., Reddy, G. B., & Reddy, V. D. (2008). Structural and kinetic properties of Bacillus subtilis S-adenosylmethionine synthetase expressed in Escherichia coli. Biochimica et Biophysica Acta, 1784, 1949–1958.
Chu, J., Qian, J., Zhuang, Y., Zhang, S., & Li, Y. (2013). Progress in the research of S-adenosylmethionine production. Applied Microbiology and Biotechnology, 97, 41–49.
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This research was supported by a grant from Seconda Università of Naples.
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Porcelli, M., Ilisso, C.P., De Leo, E. et al. Biochemical Characterization of a Thermostable Adenosylmethionine Synthetase from the Archaeon Pyrococcus Furiosus with High Catalytic Power. Appl Biochem Biotechnol 175, 2916–2933 (2015). https://doi.org/10.1007/s12010-015-1476-7
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DOI: https://doi.org/10.1007/s12010-015-1476-7