Purification of NAD+ glycohydrolase from human serum

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COŞKUN Ö., Nurten R.

Oncology Letters, cilt.6, sa.1, ss.227-231, 2013 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 6 Sayı: 1
  • Basım Tarihi: 2013
  • Doi Numarası: 10.3892/ol.2013.1335
  • Dergi Adı: Oncology Letters
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.227-231
  • Anahtar Kelimeler: ADP-ribose, Affinity chromatography, Isoelectric focusing, NAD glycohydrolase, Soluble CD38
  • İstanbul Yeni Yüzyıl Üniversitesi Adresli: Hayır

Özet

In the present study, NAD+ glycohydrolase was purified from serum samples collected from healthy individuals using ammonium sulfate fractionation, Affi-Gel blue (Cibacron Blue F3GA) affinity chromatography, Sephadex G-100 column chromatography and isoelectric focusing. The final step was followed by a second Sephadex G-100 column chromatography assay in order to remove the ampholytes from the isoelectric focusing step. In terms of enhancement of specific activity, the NAD+ glycohydrolase protein was purified ~480-fold, with a yield of 1% compared with the initial serum fraction. The purified fraction appeared to be homogeneous, with a molecular weight of 39 kDa, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, and also corresponded to the soluble (monomeric) form of surface antigen CD38.