Event Title
Purification of Actin Capping Protein Alpha Subunit Fusion Proteins
Location
CSU
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn Hart
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Actin is a cytoskeletal component that contributes to cell motility and shape. Actin is regulated by a variety of proteins including capping protein (CP). CP, composed of alpha (α) and beta (β) subunits, regulates the length and stability of actin filaments. There are two forms of the alpha subunit, αl and α2, which show 90% sequence homology across vertebrates. The regions of divergence distinguish αl and α2, and are also highly conserved. This suggests that αl and α2 have unique cellular functions. We are using a bacterial system to over-express α2 fusion protein, composed of α2 protein and maltose binding protein. The fusion protein was separated from the bacterial background using affinity chromatography, and the purity was assessed by using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis. A Bradford Protein Assay was used to quantify the concentration of the purified protein. The purified protein was used as an immunogen to generate a polyclonal antibody in chicken. In future studies, the chicken antibody, along with previously made rabbit antibody, will be used to perform double immunolocalization studies to ascertain αl and α2 specific localization in murine tissues.
Purification of Actin Capping Protein Alpha Subunit Fusion Proteins
CSU
Actin is a cytoskeletal component that contributes to cell motility and shape. Actin is regulated by a variety of proteins including capping protein (CP). CP, composed of alpha (α) and beta (β) subunits, regulates the length and stability of actin filaments. There are two forms of the alpha subunit, αl and α2, which show 90% sequence homology across vertebrates. The regions of divergence distinguish αl and α2, and are also highly conserved. This suggests that αl and α2 have unique cellular functions. We are using a bacterial system to over-express α2 fusion protein, composed of α2 protein and maltose binding protein. The fusion protein was separated from the bacterial background using affinity chromatography, and the purity was assessed by using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis. A Bradford Protein Assay was used to quantify the concentration of the purified protein. The purified protein was used as an immunogen to generate a polyclonal antibody in chicken. In future studies, the chicken antibody, along with previously made rabbit antibody, will be used to perform double immunolocalization studies to ascertain αl and α2 specific localization in murine tissues.
