Binding Analysis of the SRC-SH2 Employing a Robotic Spotting Technique and Preliminary SH2 Database Information
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Cell signaling is largely accomplished through proteins interacting in a regulated manner to produce a coherent output that controls cellular behavior. When cell signaling goes awry, it results in a plethora of disease states, including cancer, diabetes and autoimmune diseases. The SH2 domain is a protein-protein interaction motif that allows transmission of signals from receptor tyrosine kinases and acts at an early stage in many signaling pathways. Not surprisingly, proteins containing SH2 domains, or the tyrosine kinases that depend upon them, are mutated in many cancers. Therefore, studying SH2 domains is key to understanding and potentially controlling cancer. This study utilized the SPOT synthesis apparatus of producing high density peptide arrays and conducted a test run Western blot analysis of the well known Rous Sarcoma Virus oncoprotein, the SRC-SH2 domain. The results confirmed the hypothesized binding preference of the SRC-SH2 to a peptide motif of phosphotyrosine-glutamic acid-glutamic acid-isoleucine (p YEEI). Because these results verified previously known information, the test run was considered to be a success and the SPOT synthesis apparatus could then be used to uncover the binding preferences of cancer related SH2 domains. The NCBI will also be utilized in order to begin the task of compiling all the known data on SH2 domains into a database which will be shared among research institutions working with SH2s. The data collected thus far includes diseases which are directly linked to SH2 malfunction, SH2 knockouts in mice and a clarification of SH2nomenclature. The disease table especially provides the importance for studying SH2 domains where half of the disease causing SH2s directly cause cancer.