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    Öğe
    Purification of NAD+ glycohydrolase enzyme from erythrocyte membrane
    (2010) Coşkun, Özlem; Nurten, Rüstem
    Objective: In this study our object was the purification of the erythrocyte membrane bound NAD glycohydrolase of the CD38 surface antigen that is involved in the lymphocyte differentiation and activation and is also found on the erythrocyte membrane. We think that the purification of the enzyme will elucidate the structure and the activity of the CD38 and consequently the maturation, aging and death of the hemopoietic cells and the pathologic mechanisms involved in this process. Material and Methods: Fresh blood samples from the healthy individuals were obtained by anticoagulated with heparin. Solubilized eythrocyte ghosts were purified utilizing hydroxylapatite resin, Sephadex G-100 (gel filtration chromatography), Cu++ -bound IDA (iminodiacetic acid), chromatography, octyl agarose, DEAE cellulose column chromatography steps, respectively. Results: A protein band with a molecular weight 45 000 Dalton was obtained in the SDS-PAGE analysis after the purification. Conclusion: Membrane bound NAD+ glycohydrolase activity after purification was found 257 times higher than the initial erythrocyte ghost yield. The yield of the purification was 1.4%. Copyright © 2010 by Türkiye Klinikleri.
  • [ X ]
    Öğe
    The role of CD38 and CD31 in red cell interaction with endothelial cells
    (2011) Coşkun, Özlem; Varol, Başak; Nurten, Rüstem
    Objective: Abnormalities in endothelial cell structure and function may indicate the diseases such as thrombosis, atherosclerosis and vasculitis in the blood cell wall. The role of endothelial cells in these biological events was understood better with culturing. The aim of this study is to determine the pathological role of sCD38 in the process of thrombosis formation and to create a model system. Material and Methods: Human Umbilical Erythrocyte Vein Cells (HUVEC) interaction was observed with radioactive labelled erythrocytes. HUVEC-erythrocyte interaction was determined with Western Blot analysis and with HUVEC through CD31 ligand was showed with fluoresence microscopy technique. Moreover, this interaction was observed with phase contrast microscopy using Giemsa erythrocyte dye comparing the healthy individuals to atherosclerosis and cancer patients. Results: In this present study it was concluded that HUVEC-erythrocyte interaction was shown a significant enhancement in atherosclerosis and cancer patients comparing to healthy individuals by using fluoresence and phase contrast microscopy techniques. These findings were also supported with Western-Blot analysis. Conclusion: It is believed that our study will enlighten future planned clinical studies about cancer. Copyright © 2011 by Türkiye Klinikleri.