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LOCALIZING THE CYTOHESIN 2/ARNO BINDING SITE IN THE HUMAN PROTEIN INTERACTION PROTEIN FOR CYTOHESIN FACTORS (IPCEF).
Author(s) -
Richardson Clarice Leah,
Santy Lorraine C
Publication year - 2009
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.23.1_supplement.863.1
Subject(s) - mutant , microbiology and biotechnology , dna binding protein , immunoprecipitation , transfection , plasma protein binding , biology , mutagenesis , binding site , binding protein , gene , genetics , transcription factor
The study and application of locating the Cytohesin 2/ARNO binding site in the Human Protein Interaction Protein for Cytohesin Factors (IPCEF) is a very vital skill. Previous studies indicate that Cytohesin 2/ARNO binds to the IPCEF Protein in the last 121 amino acids. Consequently, our research focused on localizing the binding site of Cytohesin 2/ARNO in the IPCEF Protein while clarifying whether there are any other proteins that have the same function (binding ability) in the region where Cytohesin 2/ARNO binds in the IPCEF Protein. Next, IPCEF DNA was transformed; cultured; and Maxi and Mini prep QIAGEN kits were used to purify the IPCEF DNA. Electrophoresis; mutagenesis; digestion; co‐immunoprecipitation; transfection; and Western Blot were performed. Mutant 389 and Mutant 405 were successfully created and tested for their binding ability with Cytohesin 2/ARNO in the IPCEF Protein. Mutant 389 did not retain its binding ability while Mutant 405 retained its binding ability with Cytohesin 2/ARNO. These results are very vital because it lets us know that at least the regions from amino acid 389 (mutant 389) and amino acid 405 (mutant 405) are needed when conducting tests to locate the Cytohesin 2/ARNO binding site in the IPCEP Protein. In the future, our focus will be to further investigate where the binding site of Cytohesin 2/ARNO is located in the Human IPCEF Protein.