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Goat antibody to GST coupled to the CM5 sensor chip (BIAcore) was used for the binding of the 53BP2-GST fusion protein. The p53 core domain protein at 10, 40, 60, 80, and 100 nM concentrations was injected onto the chip in 100 mM NaCl for 2 min. followed by 2 min of dissociation phase. The binding data were analyzed by means of the BIAevaluation software.
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2-terminus, residues 491 to 495 of the n-src loop of the SH3 domain, and residues 436 to 441 in the linker between the α helices of the fourth ankyrin repeat have poor electron density and are likely to be disordered in the crystals.
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2,-terminal sequence analyses; W. Farley for help with the surface plasmon resonance experiments; M. Mayhew for helpful discussions; and R. Kenny for administrative assistance. Supported by the NIH (CA65698), the Pew Charitable Trusts, the Arnold and Mabel Beckman Foundation, the Dewitt Wallace Foundation, and the Samuel and May Rudin Foundation. Coordinates have been deposited with the Brookhaven Protein Data Bank (code 1YCS).
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