Molecular cloning expression and biochemical characterization of the human lens ALDH1 A1

Tian Lin Xiao, Ansari H. Naseem

Research output: Contribution to journalArticle

Abstract

• Aim: To clone, express and characterize the biochemical properties of recombinant human lens ALDH1A1 protein. • Methods: The complete coding sequence of ALDH1A1 from human lens library was cloned and expressed in E. coli. with PCR and plasmid transformation. Recombinant human lens ALDH1A1 protein was purified using His-tag column, and its biochemical properties such as enzyme activity, reaction buffer, cofactor, reductant and pH were studied. • Results: The coding region of human lens ALDH1A1 cDNA encodes a 1506bp DNA and a 501 amino acid protein (MW = 54. 8 kDa) that is 100% identical to human liver ALDH1A1 and shares a 85% with rat and mouse liver. The activity of recombinant human lens ALDH1A1 can be maintained for a longer period while eluted by thrombin than by imidazole, which has optimum activity with sodium pyrophosphate as a reaction buffer at pH 8. 0, preferring NAD as cofactor and dTT as reductant, and exhibits increase activity with higher pH. • Conclusion: ALDH1A1 exhibits similar characters as human liver ALDH1A1.

Original languageEnglish (US)
Pages (from-to)676-679
Number of pages4
JournalInternational Journal of Ophthalmology
Volume9
Issue number4
DOIs
StatePublished - Apr 25 2009

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Keywords

  • ALDH
  • Cataract
  • HNE
  • Lipid peroxidation

ASJC Scopus subject areas

  • Ophthalmology

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