When quantifying the binding energy of a ncAA-protein complex, we observe varying energies for each trial (noise spikes of -0.3 to 0.7 REUs for binding energy & 13 to 21 REUs for folding energy) at a given residue after mutagenesis which doesn’t seem correct given our system.
Our workflow is as follows: we separate a protein-cAA chain, “score” the constituents, mutate the AA to an ncAA & “re-score.” Do you know why we could be experiencing such fluctuations in REUs given the same complex/interface residues? In addition, is there a distinct difference between 2 & 3 or 10 & 20 REUs (same order of magnitude) or can the difference between these be considered negligible where the “trend” is most important given a residue with varying scores when mutated.
When quantifying the binding energy of a ncAA-protein complex, we observe varying energies for each trial (noise spikes of -0.3 to 0.7 REUs for binding energy & 13 to 21 REUs for folding energy) at a given residue after mutagenesis which doesn’t seem correct given our system.
Our workflow is as follows: we separate a protein-cAA chain, “score” the constituents, mutate the AA to an ncAA & “re-score.” Do you know why we could be experiencing such fluctuations in REUs given the same complex/interface residues? In addition, is there a distinct difference between 2 & 3 or 10 & 20 REUs (same order of magnitude) or can the difference between these be considered negligible where the “trend” is most important given a residue with varying scores when mutated.