Apr 20th, 1:00 PM - 4:00 PM


Development of a Recombinant Protein Expression System for the Homogenious Production of Human Carbonyl Reductase

Faculty Sponsor

Dr. Henry A. Charlier


Anthracyclines are a common and potent chemotherapy treatment for cancers of the lung, breast, uterus, and ovaries, as well as lymphomas and leukemias. One of the major side effects of anthracycline treatment however is the cumulative dose dependent cardiotoxicity. Studies suggest this cardiotoxicity is linked to the reduction of the side chain carbonyl found on this molecule. Human carbonyl reductase catalyzes the reduction of the carbonyl side chain of these molecules. This reduction leads to an anthracycline metabolite that has been linked to the development of the cardiotioxicity. Important to the ongoing studies is the development of a readily available source of human carbonyl enzyme. A clone of the gene that encodes for HCBR was used to create a gene construct that when expressed would lead to human carbonyl reductase with a 29 amino acid polyhistidine peptide attached. This poly histidine tail has a strong affinity for Ni2+ and as such provides a convenient means of purification using a single chromatography step. The resulting HCBR isolated by this method showed a decrease in catalytic activity from the native enzyme. It is believed that this interference is caused by the addition of this amino acid tail. The focus of my research currently is the shortening of this amino acid tail to include only a 6 amino acid polyhistidine peptide. It is believed that such a short peptide will not impair enzyme function. (NIH Grant # P20 RR016454)