Isolation and identification of diadenosine 5',5'''-P1,P4-tetraphosphate binding proteins using magnetic bio-panning.
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.
Pharmacy School of Shanghai Jiao Tong University, Shanghai, China.
Wei
W
Guo
M Ameruddin
MA
Azhar
Yuhong
Y
Xu
Michael
M
Wright
Ahmed
A
Kamal
Andrew D
AD
Miller
D001710:Biotin / Q000138:chemical synthesis / Q000737:chemistry; D002352:Carrier Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D015226:Dinucleoside Phosphates / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D008280:Magnetics; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry; D012015:Reference Standards
D002352:Carrier Proteins; D015226:Dinucleoside Phosphates; C020733:diadenosine tetraphosphate; D001710:Biotin