Human serum albumin is the most abundant protein in human plasma and is responsible for shuttling numerous endogenous and exogenous ligands throughout the body. Research on competitive binding to human serum albumin is of particular interest because it more accurately represents conditions encountered physiologically. Four mutations to albumin were created and expressed recombinantly using Pichia pastoris. Results of circular dichroism and fluorescence spectroscopy employed to probe the secondary and tertiary structure of recombinant albumin found incorrectly folded protein. Isothermal titration calorimetry was used to determine the binding constants of heme, ibuprofen and warfarin to albumin. An attempt to determine the competitive binding constant of warfarin binding to heme bound human serum albumin was also made using isothermal titration calorimetry. Failure to express large quantities of correctly folded recombinant protein hindered our efforts to determine binding constants for mutant human serum albumin using isothermal titration calorimetry.
|Advisor:||McClain, Brian L.|
|Commitee:||McAbee, Douglas D., Weers, Paul M. M.|
|School:||California State University, Long Beach|
|School Location:||United States -- California|
|Source:||MAI 51/03M(E), Masters Abstracts International|
|Subjects:||Analytical chemistry, Biochemistry|
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