Honors Theses
Date of Award
2005
Document Type
Undergraduate Thesis
Department
Chemistry and Biochemistry
First Advisor
Susan Pedigo
Relational Format
Dissertation/Thesis
Abstract
Cadherin is a cell adhesion protein that is significant in tissue formation. The protein is comprised of an extracellular region, a transmembrane segment and a cytoplasmic domain. The extracellular region contains five independently folded Pbarrel domains. They communicate with one another through calcium ions that bind at the interface between each domain. Our laboratory is interested in characterizing the interactions that the extracellular domains participate in. As a part of this overall goal, analytical techniques must be developed to assess the size of the constmcts. This is important for basic characterization of the recombinant proteins. Determining the change in size is also critical since these extracellular domains form higher ordered structures in vivo (dimer, tetramer, hexamer,...). Size exclusion chromatography (SEC) is a standard method for characterizing the size of proteins. The molecular weight of five cadherin constructs as determined experimentally using size exclusion chromatography (SEC) was approximately twice that of the computed value. This is misleading given the tendency for cadherin to dimerize in vivo. Alternatively, the SEC column was calibrated according to the Stokes radius of the standard proteins. Using this revised standard curve, cadherin constructs appeared to have Stokes’ radii very similar to that determined computationally for these constructs using the program HYDROPRO. Thus, calibration of SEC columns according to the Stokes’ radius showed that the cadherin constructs were monomeric with sizes very similar to that predicted.
Recommended Citation
Anazia, Gregg Oyemachi, "Size Exclusion Chromatography: Estimation of Protein Shape" (2005). Honors Theses. 2170.
https://egrove.olemiss.edu/hon_thesis/2170
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