Title
Development of a Recombinant Protein Expression System for the Production of Human Carbonyl Reductase
Document Type
Presentation
Publication Date
April 2010
Faculty Sponsor
Dr. Henry Charlier and Dr. Kristen Mitchell
Abstract
Anthracyclines are commonly used to treat a variety of cancers, but are limited in use by a dose-dependent cardiotoxicity. This cardiotoxicity is linked to the enzymatic reduction of the anthracyclines by human carbonyl reductase (HCBR). Important to the ongoing studies is the development of a readily available source of human carbonyl enzyme. Recombinant forms of HCBR were developed whereby peptides harboring a 6X-polyhistidine tail were added to either the amino- or carboxy-terminus of the protein for purification via nickel affinity chromatography. The protein with the carboxy-terminal modification had markedly reduced turnover number and catalytic efficiency. The specific activity of the amino-terminal modified enzyme is closer to the native enzyme. Preliminary results indicate HCBR is less affected by amino-terminal modification than modification at the carboxy-terminus (NIH Grant # P20 RR016454).