Purification of NAD+ glycohydrolase from human serum

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COŞKUN Ö., Nurten R.

Oncology Letters, vol.6, no.1, pp.227-231, 2013 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 6 Issue: 1
  • Publication Date: 2013
  • Doi Number: 10.3892/ol.2013.1335
  • Journal Name: Oncology Letters
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.227-231
  • Keywords: ADP-ribose, Affinity chromatography, Isoelectric focusing, NAD glycohydrolase, Soluble CD38
  • İstanbul Yeni Yüzyıl University Affiliated: No

Abstract

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.