title: [Purification of recombinant AKR7A5 protein and measurement of substrate specificity of AKR7A5 towards naphthoquinone and its derivatives].
abstract: OBJECTIVE
  To investigate the substrate specificity of mouse aldo-keto reductase AKR7A5 protein towards naphthoquinone and its derivatives.
  
  
  METHODS
  The recombinant His-tagged AKR7A5 fusion protein in E.coli BL21pLysS cell strain was induced by IPTG and purified using FPLC system through HiTrap affinity column. The purified recombinant AKR7A5 protein was confirmed by SDS-PAGE and Western blotting. AKR enzyme assay was applied to measure the substrate specificity of recombinant AKR7A5 protein towards naphthoquinone and its derivatives.
  
  
  RESULTS
  Recombinant His-AKR7A5 was successfully purified as confirmed by SDS-PAGE and Western blotting. AKR enzyme assay indicated that the recombinant AKR7A5 protein exhibited mild substrate specificity towards lawsone and low specificity towards juglone and vitamine K3,  but no activity towards 1, 4-naphthoquinone.
  
  
  CONCLUSIONS
  AKR7A5 has selective substrate specificity towards naphthoquinone derivatives,  suggesting that the aldo-keto reductase could play an important role in metabolism of certain naphthoquinone derivatives.
authors.affiliation: School of Pharmacy, First Affiliated Hospital, China Medical University, Shenyang, China. danlicmu@hotmail.com
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.forename: Ya-ru
authors.initials: YR
authors.lastname: Liu
authors.forename: Qi-shan
authors.initials: QS
authors.lastname: Zhang
authors.forename: Yang
authors.initials: Y
authors.lastname: Chu
mesh_terms: D000429:Alcohol Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009285:Naphthoquinones / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D009285:Naphthoquinones; D011993:Recombinant Fusion Proteins; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase; C000614209:AKR7A5 protein, mouse