Novel Procedure Modifications for the Growth and Purification of the Mutant Forms D34K, D79K, and E118K of Horse Heart Cytochrome c Peroxidase
Presentation Type
Poster/Portfolio
Presenter Major(s)
Biology
Mentor Information
Cory DiCarlo
Department
Chemistry
Location
Kirkhof Center KC55
Start Date
11-4-2012 9:00 AM
Keywords
Health, Life Science
Abstract
The required modifications for the growth and purification method of D34K, D79K, and E118K novel protein mutations were determined. The mutation caused changes to the growth ability of the proteins which resulted in a decreased amount of hemin needed. The hemin amount was decreased because extra hemin could disrupt the purification process and destroy the protein. Also, the amino acid's interactions with the solutions in the environment were altered due to the mutation's locations on the surface of the protein and the mutation changing the amino acid from a polar acidic to a polar basic. The change from acidic to basic caused the mutated protein to elute out with a lower concentrated mobile phase compared to the wild-type CcP. Crystallization of the D34K and D79K was achieved through the use of ion exchange chromatography, size exclusion chromatography, and dialysis.
Novel Procedure Modifications for the Growth and Purification of the Mutant Forms D34K, D79K, and E118K of Horse Heart Cytochrome c Peroxidase
Kirkhof Center KC55
The required modifications for the growth and purification method of D34K, D79K, and E118K novel protein mutations were determined. The mutation caused changes to the growth ability of the proteins which resulted in a decreased amount of hemin needed. The hemin amount was decreased because extra hemin could disrupt the purification process and destroy the protein. Also, the amino acid's interactions with the solutions in the environment were altered due to the mutation's locations on the surface of the protein and the mutation changing the amino acid from a polar acidic to a polar basic. The change from acidic to basic caused the mutated protein to elute out with a lower concentrated mobile phase compared to the wild-type CcP. Crystallization of the D34K and D79K was achieved through the use of ion exchange chromatography, size exclusion chromatography, and dialysis.