Determination of protein-bound trace elements in human cell cytosols of different organs and different pathological states.

Abstract

In the human body, there exists over 200 different cell types, which differ in size and structure and have specialised functions in the organism. Therefore it can be assumed that these different cells also contain different proteins necessary to carry out the respective specialised functions. This supposed different metalloprotein composition in different human organs cannot be demonstrated by determination of total element concentrations. Therefore investigations of the different protein-bound forms of the elements were achieved by speciation analysis: The biomolecules were separated by size exclusion chromatography and the elements detected on-line in the eluate by a hyphenated inductively coupled plasma mass spectrometer (ICP-MS). For the interpretation of the obtained element profiles, an identification of the signals and their assignment to different metalloproteins was necessary. This identification was carried out by means of specific protein assays, i.e. enzymatic assays or immunochemical reactions, in collected fractions of the chromatographic separations. A comparison of the element binding pattern in cytosols of different human organs was then possible. The optimised method was applied to tissue cytosols of different human organs. As expected, the element patterns varied for different organs of the same patient and for the same organ of patients with different diseases. Metalloproteins and their bound metals could consequently be considered as biological markers for physiological differences or pathological changes in human tissues.