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Abstract No.: TuP-216
Session: Proteomics: New Methods
Presentation date: Tue, Aug 29, 2006
Presentation time: 14:30 – 16:00

Use of Combined Orthogonal Chromatographic Techniques and Mass Spectrometry in the Characterization of Metallo-biomolecules (Metallomics)

Jose L. Gomez-Ariza1, Tamara Garcia-Barrera1, Ana Arias-Borrego1

1 Universidad de Huelva, Huelva, Spain

Correspondence address: Jose L. Gomez-Ariza, Universidad De Huelva, Quimica Y Ciencia De Los Materiales, Campus de el Carmen, Huelva, 21007 Spain.

Keywords: Chromatography, Liquid -, High Pressure (HPLC); Mass Spectrometry; Mass Spectrometry, Inductively Coupled Plasma; Metalloprotein.

Novel aspect: Metallo-biomolecules characterization through the metal.

 

The analytical characterization metallobiomolecules is crucial in many studies related to endogenous chemical species from biological systems, in which identification and/or structure characterization of these species have been required to assess their availability, assimilation/excretion, beneficial or toxic effects. This topic is today very critical due to the increasing interest for the systematic acquisition of information related to proteins and other biomolecules involved in cell function.1 Special attention is paid on metals in metalloproteins (e.g., metalloenzymes, metal transporters, metal probes, and others), and on others non-metallic elements involved in the cell machinery, such as sulphur in metallothioneins, or phosphorous in cell signalling phosphoproteins. Small size metallomolecules are also important in the overall cell metabolism and function,1,2 whose complete characterization requires the study of both high (HMW) and low molecular weight (LMW) molecules.

For this purpose multidimensional analytical approaches in which a separation step based on orthogonal separation techniques are combined in parallel with an atomic detector (namely, ICP-MS) and some other techniques for molecular structure, mainly tandem mass spectrometry. This new approach has been called metallomics.3,4

A generic metallomics analytical approach (MMA), with special attention to metalloproteins, has been proposed,4 which consists of three main components: (i) a separation technique (e.g. HPLC, CE, 2DE) for target species isolation from the matrix and their time-resolved introduction into the detector— discrimination component; (ii) a high-sensitivity element detector such as ICP-MS, for element quantification— element-tag component; and (iii) an instrumental system for known molecule characterization, based on mass spectrometry, especially tandem MS— structural component. This approach has been used for environmental, food and biomedicine issues.

1. J. Szpunar, Analyst 130, 442 (2005).
2. C. E. Outten and T. V. O’Halloran, Science 292, 2488 (2001).
3. J. Szpunar, Anal. Bioanal. Chem. 378, 54 (2004).
4. J. L. Gomez-Ariza, T. Garcia-Barrera, F. Lorenzo, V. Bernal, M. J. Villegas and V. Oliveira, Anal. Chim. Acta 524, 15 (2004).