Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/16210
Title: Development and application of mass spectrometry based proteomics technologies to decipher ku70 functions
Authors: MENG WEI
Keywords: Ku70; Mass spectrometry; TAP purification; acetylation; Empore disk extraction; PTM
Issue Date: 25-May-2007
Source: MENG WEI (2007-05-25). Development and application of mass spectrometry based proteomics technologies to decipher ku70 functions. ScholarBank@NUS Repository.
Abstract: Ku70 is a protein with multiple biological functions. Given the dual role of Ku70 in DNA repair and apoptosis, it might be a therapeutic target for cancer treatment. We employed epitope-tag purification and mass spectrometry based proteomic methods to characterize the Ku70 protein complex to unravel its biological functions. We also developed a novel effective peptide purification and concentration method for in-gel sample. The pulled-down complex was analyzed by both SDS-PAGE coupled to MALDI-TOF/TOF-MS and shotgun LC-MS/MS proteomics approaches. Consequently, 151 proteins were characterized in Ku70 protein complexes. These proteins play a diverse range of biological functions from DNA repair to transcriptional regulation to cellular signal mediator. As Ku70 acetylation has been reported to be a pivotal post translational modification that regulates Ku70 activities, we finally identified the potential acetylation sites of Ku70 by both in vivo and in vitro acetylation analysis.
URI: http://scholarbank.nus.edu.sg/handle/10635/16210
Appears in Collections:Master's Theses (Open)

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