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Isolation and identification of diadenosine 5',5'-P1,P4-tetraphosphate binding proteins using magnetic bio-panning
Authors:Guo Wei  Azhar M Ameruddin  Xu Yuhong  Wright Michael  Kamal Ahmed  Miller Andrew D
Institution:Pharmacy School of Shanghai Jiao Tong University, Shanghai, China.
Abstract:We report the development of a synthetic, biotin-conjugated diadenosine tetraphosphate (Ap(4)A)-'molecular hook' attached to magnetic beads enabling the isolation of Ap(4)A-binding proteins from bacterial cells or mammalian tissue lysates. Characterisation and identification of isolated binding proteins is performed sequentially by mass spectrometry. The observation of positive controls suggests that these newly observed proteins are putative Ap(4)A-binding partners, and we have expectations that others can be found with further technical improvements in our methods.
Keywords:Ap4A  diadenosine tetraphosphate  ATP  adenosine 5′-triphosphate  ESI-MS  electrospray ionisation mass spectroscopy  IMP  inosine 5′-monophosphate  IMPDH  IMP dehydrogenase  NAD  nicotinamide adenine dinucleotide  NpnN  dinucleoside polyphosphate
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