Cytosolic proteins from neutrophilic granulocytes: A comparison between patients with severe chronic neutropenia and healthy donors

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Abstract
A modified technique of two-dimensional gel electrophoresis (2-DE) was used to investigate differences in the pattern of cytosolic proteins of neutrophilic granulocytes from patients with severe congenital neutropenia, cyclic neutropenia, and idiopathic neutropenia in comparison with healthy donors. At the time of study, all patients tested received treatment with a recombinant human granulocyte colony-stimulating factor (r-metHuG-CSF; Filgrastim, Amgen). Using the Investigator 2-D Electrophoresis System™ (Millipore) we were able to detect more than 1000 protein spots in the cytosol of neutrophilic granulocytes from both patients and healthy controls. We investigated six patients with severe congenital neutropenia, five patients with cyclic neutropenia, four patients with idiopathic neutropenia, and 13 healthy donors. In the cytosol of neutrophilic granulocytes from patients we found an additional protein spot. This protein spot (molecular mass approximately 32.4 kDa, pI about 5.5) could be identified by internal sequencing after in-gel digestion with endoproteinase Lys-C as tropomyosin. The importance of the overexpression of tropomyosin in neutrophilic granulocytes from patients with severe chronic neutropenia is not yet understood.

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