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We believe our estimates of alignment overextension (7 10% of alignments) are conservative, both because sequence databases are growing, allowing similarity searches to identify closer homologs, and because many proteins comprise multiple domains.
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Note that such an estimate of the alignment error rate may be underestimated because wrongly aligned human reads tend to be related to repetitive sequences in human and to be mapped back to the human sequences.
From an initial estimate of the alignment, a new similarity matrix is generated using the relative alpha carbon coordinates that result from a multi-body superposition.
Prior to the start of the experiment, we obtained an estimate of subjects' alignment and bisection thresholds at each separation using an adaptive staircase procedure, with a total of 50 presentations.
To estimate the effect of alignment ambiguity, we analyzed the sites remaining after exclusion of the RRM.
Thus PSI-BLAST-based profile searches against consensus sequences might incorrectly estimate statistical significance of alignment scores.
We can calculate the reference alignment probability P ref(A) in Equation (1) by randomly sampling a set of protein pairs, each with the same lengths as the sequence S and template T, respectively, and then estimate the probability of alignment A based on these randomly sampled protein pairs.
For estimating the effect of alignment shifts in models with otherwise correct fold and stereochemistry, we created pseudomodels starting from the original protein structure and introducing threading errors of increasing magnitude for different representative structure architectures from CATH (Cuff et al., 2011).
This procedure yields estimates of mechanical axis alignment similar to full-limb weight-bearing radiographs (R=0.54) but without exposure to radiation [ 31].
Importantly, MSAprobs has the further advantage of providing, for each column of an alignment, dependable estimates of the confidence of the alignment at the site.
Bootstrap supports were estimated from 1,000 samples of alignment generated by the SEQBOOT program of PHYLIP v3.69 (Felsenstein 1989).
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