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Structural Analysis of Cortactin After Anion Exchange Chromatography
Author(s) -
Graves Gabriel,
Kruchten Anne
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.792.25
Subject(s) - cortactin , in vitro , elution , in vivo , cytoskeleton , ion chromatography , polyacrylamide gel electrophoresis , affinity chromatography , chemistry , microbiology and biotechnology , staining , silver stain , ion exchange , chromatography , biochemistry , biophysics , biology , cell , ion , genetics , organic chemistry , enzyme
Cortactin is a cytoskeletal actin‐binding protein that is upregulated in several types of metastatic cancers. Further research on its three‐dimensional structure may allow for greater understanding of its role in cancers. Purification of our in vitro expressed His‐tagged cortactin from E. coli was done through affinity purification with Ni2+ resin followed by anion exchange chromatography. Elution fractions of the protein were analyzed via polyacrylamide gels both in native and SDS environments to analyze both length and hydrodynamic size of the protein. Silver staining of subsequent gels allowed for precise analysis of banding pattern and relative strength. Our results demonstrate variations in banding patterns with increasing concentration of KCl elution on native‐PAGE, suggesting cortactin has a variable three dimensional structure in vitro. Future work will determine if these structural differences are maintained in vivo . Support or Funding Information This work is supported by the College of Saint Scholastica SSC Summer Research Fellowships. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .