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 language | English (US) |
|---|---|
| Pages (from-to) | 676-679 |
| Number of pages | 4 |
| Journal | International Journal of Ophthalmology |
| Volume | 9 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 25 2009 |
| Externally published | Yes |
Keywords
- ALDH
- Cataract
- HNE
- Lipid peroxidation
ASJC Scopus subject areas
- Ophthalmology