Semester

Fall

Date of Graduation

2004

Document Type

Thesis

Degree Type

MS

College

School of Pharmacy

Department

Pharmaceutical Sciences

Committee Chair

Robert L. Haining.

Abstract

To date, knowledge in characterization of CYP proteins has been limited to immunoblotting, RT-PCR, immunohistochemistry and so forth; therefore, it is the intention of this study to investigate: (1) What kind of CYP2D protein is present in brain with modern proteomic tools; (2) Whether, CYP2D6 in brain is same as the liver CYP2D6; and (3) If CYP2D7 is present, can CYP2D6 polymorphism be found in the brain?;To answer these questions, this study used a one-step method of isolation of protein by immunoprecipitation followed by its identification using 2D-blots, nanospray LCMS, immunoblots and immunohistochemistry. Probing of immunoprecipitated proteins with polyclonal CYP2D6 antibody revealed two major CYP2D6 immunoreactive bands. Similar banding pattern resulted from matched human liver microsomes suggesting there are more than one CYP2D isoform(s) exist in these tissues.

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