engleski

Purification and MALDI-MS characterization of a stress - associated glycoprotein from sera of professional soldiers

Glycoconjugates have a whole spectrum of biological roles, from those that appear trivial, to those that are crucial. Results accumulated in the past years indicate they might also play an important role in the response to stress, a complex physiological response of human organism to various threats. We have recently identified Stressin, a human serum glycoprotein, which was found to be increased in stress. Here we report purification of Stressin from sera of professional soldiers and partial characterization of its protein and carbohydrate parts using lectin blotting and Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS). Stressin was purified using combination of ammonium sulfate precipitation, ion exchange, preparative electrophoresis and reverse-phase HPLC. It was found to be highly glycosylated protein. Only 21, 9 kDa (out of 36, 7 kDa) was the protein part, whereas remaining 40% of the mass originated from N-linked oligosaccharides. The carbohydrate part contained 12 sialic acids moieties, nearly 90% of which were lost due to post-source decay in the field-free tube.

psychological stress; human serum glycoprotein; protein purification; MALDI-MS; glycosylation; lectins

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