| Title |
Purification and Mechanism of Human Aldehyde Oxidase Expressed in Escherichia coli
|
|---|---|
| Published in |
Drug Metabolism and Disposition, September 2009
|
| DOI | 10.1124/dmd.109.029520 |
| Pubmed ID | |
| Authors |
Joshua F. Alfaro, Carolyn A. Joswig-Jones, Wenyun Ouyang, Joseph Nichols, Gregory J. Crouch, Jeffrey P. Jones |
| Abstract |
Human aldehyde oxidase 1 (AOX1) has been subcloned into a vector suitable for expression in Escherichia coli, and the protein has been expressed. The resulting protein is active, with sulfur being incorporated in the molybdopterin cofactor. Expression levels are modest, but 1 liter of cells supplies enough protein for both biochemical and kinetic characterization. Partial purification is achieved by nickel affinity chromatography through the addition of six histidines to the amino-terminal end of the protein. Kinetic analysis, including kinetic isotope effects and comparison with xanthine oxidase, reveal similar mechanisms, with some subtle differences. This expression system will allow for the interrogation of human aldehyde oxidase structure/function relationships by site-directed mutagenesis and provide protein for characterizing the role of AOX1 in drug metabolism. |
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Mendeley demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 3 | 5% |
| Germany | 1 | 2% |
| Unknown | 59 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 13 | 21% |
| Student > Ph. D. Student | 12 | 19% |
| Student > Bachelor | 7 | 11% |
| Student > Doctoral Student | 6 | 10% |
| Student > Master | 6 | 10% |
| Other | 10 | 16% |
| Unknown | 9 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 15 | 24% |
| Biochemistry, Genetics and Molecular Biology | 10 | 16% |
| Medicine and Dentistry | 9 | 14% |
| Chemistry | 9 | 14% |
| Pharmacology, Toxicology and Pharmaceutical Science | 8 | 13% |
| Other | 2 | 3% |
| Unknown | 10 | 16% |