Abstract
In contrast to the abundance of systems-oriented approaches describing changes on the transcriptome or proteome level, relatively few studies have employed the metabolome. The goal of the presented research was to identify as many intracellular metabolites as possible in a Spiroplasma melliferum extract by flow injection time-of-flight mass spectrometry. The Mollicutes class bacterium S. melliferum is a member of a unique category of bacteria that have in common the absence of a cell wall, a reduced genome, and simplified metabolic pathways. Metabolite identification was confirmed by fragmentation of previously detected ions by target mass spectrometry. The selected liquid chromatography approach, hydrophilic interaction chromatography with amino and silica columns, effectively separates highly polar cellular metabolites prior to their detection on a high accuracy mass spectrometer in positive and negative acquisition mode for each column. Here we present reliable measurement of 76 metabolites, including components of sugar, amino acid, and nucleotide metabolism. We have identified about a third of the possible intracellular S. melliferum metabolites predicted by genome annotation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- FTICR:
-
Fourier transform ion cyclotron resonance
- HILIC:
-
hydrophilic interaction liquid chromatography
- Q:
-
quadrupole detector
- QQQ:
-
triple quadrupole detector
- TOF:
-
time-of-flight
References
Rogers, F. B. (1963) Bull. Med. Libr. Assoc., 51, 114–116.
Griffiths, W. J., Karu, K., Hornshaw, M., Woffendin, G., and Wang, Y. (2007) Eur. J. Mass Spectrom. (Chichester, Eng.), 13, 45–50.
Lewis, I. A., Schommer, S. C., Hodis, B., Robb, K. A., Tonelli, M., Westler, W. M., Sussman, M. R., and Markley, J. L. (2007) Anal. Chem., 79, 9385–9390.
Want, E. J., Cravatt, B. F., and Siuzdak, G. (2005) Chembiochem, 6, 1941–1951.
Kimball, E., and Rabinowitz, J. D. (2006) Anal. Biochem., 358, 273–280.
Rabinowitz, J. D., and Kimball, E. (2007) Anal. Chem., 79, 6167–6173.
Want, E. J., Nordstrom, A., Morita, H., and Siuzdak, G. (2007) J. Proteome Res., 6, 459–468.
Monton, M. R., and Soga, T. (2007) J. Chromatogr. A, 1168, 237–246; discussion 236.
Bajad, S. U., Lu, W., Kimball, E. H., Yuan, J., Peterson, C., and Rabinowitz, J. D. (2006) J. Chromatogr. A, 1125, 76–88.
Guillarme, D., Nguyen, D. T., Rudaz, S., and Veuthey, J. L. (2007) J. Chromatogr. A, 1149, 20–29.
Wilson, I. D., Nicholson, J. K., Castro-Perez, J., Granger, J. H., Johnson, K. A., Smith, B. W., and Plumb, R. S. (2005) J. Proteome Res., 4, 591–598.
Nguyen, D. T., Guillarme, D., Rudaz, S., and Veuthey, J. L. (2006) J. Sep. Sci., 29, 1836–1848.
Walles, M., Gauvin, C., Morin, P. E., Panetta, R., and Ducharme, J. (2007) J. Sep. Sci., 30, 1191–1199.
Cai, S. S., Short, L. C., Syage, J. A., Potvin, M., and Curtis, J. M. (2007) J. Chromatogr. A, 1173, 88–97.
Lacorte, S., and Fernandez-Alba, A. R. (2006) Mass Spectrom. Rev., 25, 866–880.
Hu, Q., Noll, R. J., Li, H., Makarov, A., Hardman, M., and Graham Cooks, R. (2005) J. Mass Spectrom., 40, 430–443.
Glish, G. L., and Burinsky, D. J. (2008) J. Am. Soc. Mass Spectrom., 19, 161–172.
Meeus, I., Vercruysse, V., and Smagghe, G. (2011) J. Invertebr. Pathol., 109, 172–174.
Fisunov, G. Y., Alexeev, D. G., Bazaleev, N. A., Ladygina, V. G., Galyamina, M. A., Kondratov, I. G., Zhukova, N. A., Serebryakova, M. V., Demina, I. A., and Govorun, V. M. (2011) PLoS One, 6, e21964.
Kamashev, D., Oberto, J., Serebryakova, M., Gorbachev, A., Zhukova, Y., Levitskii, S., Mazur, A. K., and Govorun, V. (2011) Biochemistry, 50, 8692–8702.
Levitskiy, S. A., Sycheva, A. M., Kharlampieva, D. D., Oberto, J., Kamashev, D. E., Serebryakova, M. V., Moshkovskii, S. A., Lazarev, V. N., and Govorun, V. M. (2011) Biochimie, 93, 1102–1109.
Alexeev, D., Kostrjukova, E., Aliper, A., Popenko, A., Bazaleev, N., Tyakht, A., Selezneva, O., Akopian, T., Prichodko, E., Kondratov, I., Chukin, M., Demina, I., Galyamina, M., Kamashev, D., Vanyushkina, A., Ladygina, V., Levitskii, S., Lazarev, V., and Govorun, V. (2011) J. Proteome Res., 11, 224–236.
Maharjan, R. P., and Ferenci, T. (2003) Anal. Biochem., 313, 145–154.
Melamud, E., Vastag, L., and Rabinowitz, J. D. (2010) Anal. Chem., 82, 9818–9826.
Kanehisa, M. (1997) Trends Genet., 13, 375–376.
Tredwell, G. D., Edwards-Jones, B., Leak, D. J., and Bundy, J. G. (2011) PLoS One, 6, e16286.
R_Development_Core_Team (2005) Vienna, Austria.
Smith, C. A., Want, E. J., O’Maille, G., Abagyan, R., and Siuzdak, G. (2006) Anal. Chem., 78, 779–787.
Kuhl, C., Tautenhahn, R., Bottcher, C., Larson, T. R., and Neumann, S. (2012) Anal. Chem., 84, 283–289.
Smith, C. A., O’Maille, G., Want, E. J., Qin, C., Trauger, S. A., Brandon, T. R., Custodio, D. E., Abagyan, R., and Siuzdak, G. (2005) Ther. Drug Monit., 27, 747–751.
Villas-Boas, S. G., Hojer-Pedersen, J., Akesson, M., Smedsgaard, J., and Nielsen, J. (2005) Yeast, 22, 1155–1169.
Lebedev, A. T. (2003) Mass-spectrometry in Organic Chemistry [in Russian], BINOM, Laboratoriya Znanii, Moscow.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © A. A. Vanyushkina, D. E. Kamashev, I. A. Altukhov, V. M. Govorun, 2012, published in Biokhimiya, 2012, Vol. 77, No. 8, pp. 1052–1066. Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM12-085, June 17, 2012.
Rights and permissions
About this article
Cite this article
Vanyushkina, A.A., Kamashev, D.E., Altukhov, I.A. et al. Identification of intracellular Spiroplasma melliferum metabolites by the HPLC-MS method. Biochemistry Moscow 77, 864–877 (2012). https://doi.org/10.1134/S000629791208007X
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S000629791208007X