Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT Agrobacterium tumefaciens requires phosphatidylcholine (PC) in its membranes for plant infection. The phospholipid N -methyltransferase PmtA catalyzes all three transmethylation reactions of phosphatidylethanolamine (PE) to PC via the intermediates monomethylphosphatidylethanolamine (MMPE) and dimethylphosphatidylethanolamine (DMPE). The enzyme uses S -adenosylmethionine (SAM) as the methyl donor, converting it to S -adenosylhomocysteine (SAH). Little is known about the activity of bacterial Pmt enzymes, since PC biosynthesis in prokaryotes is rare. In this article, we present the purification and in vitro characterization of A. tumefaciens PmtA, which is a monomeric protein. It binds to PE, the intermediates MMPE and DMPE, the end product PC, and phosphatidylglycerol (PG) and phosphatidylinositol. Binding of the phospholipid substrates precedes binding of SAM. We used a coupled in vitro assay system to demonstrate the enzymatic activity of PmtA and to show that PmtA is inhibited by the end products PC and SAH and the antibiotic sinefungin. The presence of PG stimulates PmtA activity. Our study provides insights into the catalysis and control of a bacterial phospholipid N -methyltransferase.

Bibliography

Aktas, M., & Narberhaus, F. (2009). In Vitro Characterization of the Enzyme Properties of the Phospholipid N -Methyltransferase PmtA from Agrobacterium tumefaciens. Journal of Bacteriology, 191(7), 2033–2041.

Authors 2
  1. Meriyem Aktas (first)
  2. Franz Narberhaus (additional)
References 44 Referenced 28
  1. Arondel, V., C. Benning, and C. R. Somerville. 1993. Isolation and functional expression in Escherichia coli of a gene encoding phosphatidylethanolamine methyltransferase (EC 2.1.1.17) from Rhodobacter sphaeroides. J. Biol. Chem. 268 : 16002-16008. (10.1016/S0021-9258(18)82350-X) / J. Biol. Chem. (1993)
  2. 10.1139/o59-099
  3. 10.1128/AAC.3.1.49
  4. Carman, G. M., R. A. Deems, and E. A. Dennis. 1995. Lipid signaling enzymes and surface dilution kinetics. J. Biol. Chem. 270 : 18711-18714. (10.1074/jbc.270.32.18711) / J. Biol. Chem. (1995)
  5. Cho, W., and R. V. Stahelin. 2005. Membrane-protein interactions in cell signaling and membrane trafficking. Annu. Rev. Biophys. Biomol. Struct. 34 : 119-151. (10.1146/annurev.biophys.33.110502.133337) / Annu. Rev. Biophys. Biomol. Struct. (2005)
  6. 10.1128/JB.188.5.1929-1934.2006
  7. Conde-Alvarez, R., M. J. Grilló, S. P. Salcedo, M. J. de Miguel, E. Fugier, J. P. Gorvel, I. Moriyón, and M. Iriarte. 2006. Synthesis of phosphatidylcholine, a typical eukaryotic phospholipid, is necessary for full virulence of the intracellular bacterial parasite Brucella abortus. Cell. Microbiol. 8 : 1322-1335. (10.1111/j.1462-5822.2006.00712.x) / Cell. Microbiol. (2006)
  8. Conover, G. M., F. Martínez-Morales, M. I. Heidtman, Z. Q. Luo, M. Tang, C. Chen, O. Geiger, and R. R. Isberg. 2008. Phosphatidylcholine synthesis is required for optimal function of Legionella pneumophila virulence determinants. Cell. Microbiol. 10 : 514-528. / Cell. Microbiol. (2008)
  9. Cui, Z., J. E. Vance, M. H. Chen, D. R. Voelker, and D. E. Vance. 1993. Cloning and expression of a novel phosphatidylethanolamine N-methyltransferase. A specific biochemical and cytological marker for a unique membrane fraction in rat liver. J. Biol. Chem. 268 : 16655-16663. (10.1016/S0021-9258(19)85468-6) / J. Biol. Chem. (1993)
  10. de Rudder, K. E., I. M. López-Lara, and O. Geiger. 2000. Inactivation of the gene for phospholipid N-methyltransferase in Sinorhizobium meliloti: phosphatidylcholine is required for normal growth. Mol. Microbiol. 37 : 763-772. (10.1046/j.1365-2958.2000.02032.x) / Mol. Microbiol. (2000)
  11. Dorgan, K. M., W. L. Wooderchak, D. P. Wynn, E. L. Karschner, J. F. Alfaro, Y. Cui, Z. S. Zhou, and J. M. Hevel. 2006. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases. Anal. Biochem. 350 : 249-255. (10.1016/j.ab.2006.01.004) / Anal. Biochem. (2006)
  12. Dowler, S., G. Kular, and D. R. Alessi. 2002. Protein lipid overlay assay. Sci. STKE 2002(129): PL6. / Sci. STKE (2002)
  13. Gaynor, P. M., and G. M. Carman. 1990. Phosphatidylethanolamine methyltransferase and phospholipid methyltransferase activities from Saccharomyces cerevisiae. Enzymological and kinetic properties. Biochim. Biophys. Acta 1045 : 156-163. (10.1016/0005-2760(90)90145-N) / Biochim. Biophys. Acta (1990)
  14. Gibson, K. D., J. D. Wilson, and S. Udenfriend. 1961. The enzymatic conversion of phospholipid ethanolamine to phospholipid choline in rat liver. J. Biol. Chem. 236 : 673-679. (10.1016/S0021-9258(18)64287-5) / J. Biol. Chem. (1961)
  15. Goldfine, H. 1984. Bacterial membranes and lipid packing theory. J. Lipid Res. 25 : 1501-1507. (10.1016/S0022-2275(20)34423-0) / J. Lipid Res. (1984)
  16. Hacker, S., J. Gödeke, A. Lindemann, S. Mesa, G. Pessi, and F. Narberhaus. 2008. Global consequences of phosphatidylcholine reduction in Bradyrhizobium japonicum. Mol. Genet. Genomics 280 : 59-72. (10.1007/s00438-008-0345-2) / Mol. Genet. Genomics (2008)
  17. 10.1128/JB.01423-07
  18. Haydock, S. F., J. A. Dowson, N. Dhillon, G. A. Roberts, J. Cortes, and P. F. Leadlay. 1991. Cloning and sequence analysis of genes involved in erythromycin biosynthesis in Saccharopolyspora erythraea: sequence similarities between EryG and a family of S-adenosylmethionine-dependent methyltransferases. Mol. Gen. Genet. 230 : 120-128. (10.1007/BF00290659) / Mol. Gen. Genet. (1991)
  19. Ingrosso, D., A. V. Fowler, J. Bleibaum, and S. Clarke. 1989. Sequence of the d-aspartyl/l-isoaspartyl protein methyltransferase from human erythrocytes. Common sequence motifs for protein, DNA, RNA, and small molecule S-adenosylmethionine-dependent methyltransferases. J. Biol. Chem. 264 : 20131-20139. (10.1016/S0021-9258(19)47228-1) / J. Biol. Chem. (1989)
  20. Kaneshiro, T., and J. H. Law. 1964. Phosphatidylcholine synthesis in Agrobacterium tumefaciens. I. Purification and properties of a phosphatidylethanolamine N-methyltransferase. J. Biol. Chem. 239 : 1705-1713. (10.1016/S0021-9258(18)91245-7) / J. Biol. Chem. (1964)
  21. Kanipes, M. I., and S. A. Henry. 1997. The phospholipid methyltransferases in yeast. Biochim. Biophys. Acta 1348 : 134-141. (10.1016/S0005-2760(97)00121-5) / Biochim. Biophys. Acta (1997)
  22. Kanipes, M. I., J. E. Hill, and S. A. Henry. 1998. The Schizosaccharomyces pombe cho1 + gene encodes a phospholipid methyltransferase. Genetics 150 : 553-562. (10.1093/genetics/150.2.553) / Genetics (1998)
  23. 10.1128/JB.184.15.4114-4123.2002
  24. 10.1128/JB.01183-08
  25. Kodaki, T., and S. Yamashita. 1987. Yeast phosphatidylethanolamine methylation pathway. Cloning and characterization of two distinct methyltransferase genes. J. Biol. Chem. 262 : 15428-15435. (10.1016/S0021-9258(18)47744-7) / J. Biol. Chem. (1987)
  26. López-Lara, I. M., and O. Geiger. 2001. Novel pathway for phosphatidylcholine biosynthesis in bacteria associated with eukaryotes. J. Biotechnol. 91 : 211-221. (10.1016/S0168-1656(01)00331-5) / J. Biotechnol. (2001)
  27. López-Lara, I. M., C. Sohlenkamp, and O. Geiger. 2003. Membrane lipids in plant-associated bacteria: their biosyntheses and possible functions. Mol. Plant-Microbe Interact. 16 : 567-579. (10.1094/MPMI.2003.16.7.567) / Mol. Plant-Microbe Interact. (2003)
  28. Martin, J. L., and F. M. McMillan. 2002. SAM (dependent) I AM: the S-adenosylmethionine-dependent methyltransferase fold. Curr. Opin. Struct. Biol. 12 : 783-793. (10.1016/S0959-440X(02)00391-3) / Curr. Opin. Struct. Biol. (2002)
  29. Martínez-Morales, F., M. Schobert, I. M. López-Lara, and O. Geiger. 2003. Pathways for phosphatidylcholine biosynthesis in bacteria. Microbiology 149 : 3461-3471. (10.1099/mic.0.26522-0) / Microbiology (2003)
  30. Minder, A. C., K. E. de Rudder, F. Narberhaus, H. M. Fischer, H. Hennecke, and O. Geiger. 2001. Phosphatidylcholine levels in Bradyrhizobium japonicum membranes are critical for an efficient symbiosis with the soybean host plant. Mol. Microbiol. 39 : 1186-1198. (10.1111/j.1365-2958.2001.02325.x) / Mol. Microbiol. (2001)
  31. Pessi, G., G. Kociubinski, and C. B. Mamoun. 2004. A pathway for phosphatidylcholine biosynthesis in Plasmodium falciparum involving phosphoethanolamine methylation. Proc. Natl. Acad. Sci. USA 101 : 6206-6211. (10.1073/pnas.0307742101) / Proc. Natl. Acad. Sci. USA (2004)
  32. Ridgway, N. D., and D. E. Vance. 1988. Kinetic mechanism of phosphatidylethanolamine N-methyltransferase. J. Biol. Chem. 263 : 16864-16871. (10.1016/S0021-9258(18)37471-4) / J. Biol. Chem. (1988)
  33. Ridgway, N. D., and D. E. Vance. 1987. Purification of phosphatidylethanolamine N-methyltransferase from rat liver. J. Biol. Chem. 262 : 17231-17239. (10.1016/S0021-9258(18)45514-7) / J. Biol. Chem. (1987)
  34. Sherr, S. I., and J. H. Law. 1965. Phosphatidylcholine synthesis in Agrobacterium tumefaciens. II. Uptake and utilization of choline. J. Biol. Chem. 240 : 3760-3765. (10.1016/S0021-9258(18)97107-3) / J. Biol. Chem. (1965)
  35. Shields, D. J., J. Y. Altarejos, X. Wang, L. B. Agellon, and D. E. Vance. 2003. Molecular dissection of the S-adenosylmethionine-binding site of phosphatidylethanolamine N-methyltransferase. J. Biol. Chem. 278 : 35826-35836. (10.1074/jbc.M306308200) / J. Biol. Chem. (2003)
  36. Shields, D. J., S. Lingrell, L. B. Agellon, J. T. Brosnan, and D. E. Vance. 2005. Localization-independent regulation of homocysteine secretion by phosphatidylethanolamine N-methyltransferase. J. Biol. Chem. 280 : 27339-27344. (10.1074/jbc.M504658200) / J. Biol. Chem. (2005)
  37. Sohlenkamp, C., I. M. López-Lara, and O. Geiger. 2003. Biosynthesis of phosphatidylcholine in bacteria. Prog. Lipid Res. 42 : 115-162. (10.1016/S0163-7827(02)00050-4) / Prog. Lipid Res. (2003)
  38. Tahara, Y., Y. Ogawa, T. Sakakibara, and Y. Yamada. 1987. Purification and characterization of phosphatidylethanolamine N-methyltransferase from Zymomonas mobilis. Agric. Biol. Chem. 51 : 1425-1430. / Agric. Biol. Chem. (1987)
  39. Vance, D. E., Z. Li, and R. L. Jacobs. 2007. Hepatic phosphatidylethanolamine N-methyltransferase, unexpected roles in animal biochemistry and physiology. J. Biol. Chem. 282 : 33237-33241. (10.1074/jbc.R700028200) / J. Biol. Chem. (2007)
  40. Vance, J. E. 1990. Phospholipid synthesis in a membrane fraction associated with mitochondria. J. Biol. Chem. 265 : 7248-7256. (10.1016/S0021-9258(19)39106-9) / J. Biol. Chem. (1990)
  41. Walkey, C. J., L. R. Donohue, R. Bronson, L. B. Agellon, and D. E. Vance. 1997. Disruption of the murine gene encoding phosphatidylethanolamine N-methyltransferase. Proc. Natl. Acad. Sci. USA 94 : 12880-12885. (10.1073/pnas.94.24.12880) / Proc. Natl. Acad. Sci. USA (1997)
  42. Wessel, M., S. Klüsener, J. Gödeke, C. Fritz, S. Hacker, and F. Narberhaus. 2006. Virulence of Agrobacterium tumefaciens requires phosphatidylcholine in the bacterial membrane. Mol. Microbiol. 62 : 906-9015. (10.1111/j.1365-2958.2006.05425.x) / Mol. Microbiol. (2006)
  43. 10.1128/JB.184.17.4792-4799.2002
  44. Zhu, Y., C. Qi, W. Q. Cao, A. V. Yeldandi, M. S. Rao, and J. K. Reddy. 2001. Cloning and characterization of PIMT, a protein with a methyltransferase domain, which interacts with and enhances nuclear receptor coactivator PRIP function. Proc. Natl. Acad. Sci. USA 98 : 10380-10385. (10.1073/pnas.181347498) / Proc. Natl. Acad. Sci. USA (2001)
Dates
Type When
Created 16 years, 7 months ago (Jan. 30, 2009, 8:45 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:11 p.m.)
Indexed 3 months ago (May 28, 2025, 6:05 p.m.)
Issued 16 years, 5 months ago (April 1, 2009)
Published 16 years, 5 months ago (April 1, 2009)
Published Print 16 years, 5 months ago (April 1, 2009)
Funders 0

None

@article{Aktas_2009, title={In Vitro Characterization of the Enzyme Properties of the Phospholipid N -Methyltransferase PmtA from Agrobacterium tumefaciens}, volume={191}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.01591-08}, DOI={10.1128/jb.01591-08}, number={7}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Aktas, Meriyem and Narberhaus, Franz}, year={2009}, month=apr, pages={2033–2041} }