According to the profiles of standard marker proteins such as aprotinin (MW: 6,512 Da), RNaseA (MW: 13,700 Da), albumin egg (MW: 45,000 Da) and BSA (MW: 67,000 Da) on the same column and under the same buffer conditions, these two peaks correspond to the dimer and monomer of nsp4C, respectively.
Both tetramers and hexamers bind the target peptide from p53 with retained stoichiometry of one peptide per S100B monomer, and with high affinity (lgK = 7.3±0.2 and 7.2±0.2, respectively in 10 mM BisTris, 5 mM CaCl2, pH 7.0), which is less than one order of magnitude reduced compared to dimer under the same buffer conditions.
The FEC were obtained under the same buffer conditions as our XPD helicase measurements.
To replicate the conditions used in the CD we performed the experiments under the same buffer conditions and DNA concentration.
NMR experiments were performed under the same buffer conditions but at 15 °C, with lipid vesicles composed of 15% DOPS, 25% DOPE, and 60% DOPC.
Isothermal Titration Calorimetry The heat of interaction was measured on an ITC200 microcalorimeter (Microcal), with a cell volume of 200 μL, under the same buffer conditions (20 mM Tris, pH 7.5, containing 6 mM MgCl2 and 5 mM NaCl) at 30°C. 10 3.8 μl aliquots of WT or mutant Ssa1 protein at 30 μM were injected into 350 μM of AMP-PNP.
The sample was incubated overnight in the same buffer under agitation at 4°C.
Purified ovalbumin and pure DM were run in the same buffer under the same conditions.
After washing twice in a buffer containing 50 m M Tris-HCl, 5 m M CaCl2 and 0,1% Triton X-100 (pH 7.8), the gels were incubated overnight at 37°C in the same buffer under gentle agitation.
100% survival was defined as total survival of bacteria in the same buffer and under the same conditions as in the absence of peptide.
The experiments performed under non-reducing conditions were performed in the same buffer, in the absence of DTT.
