Research
Neutral genetic drift can alter promiscuous protein functions, potentially aiding functional evolution
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* Corresponding author: Frances H Arnold frances@cheme.caltech.edu
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, 91125, USA
Biology Direct 2007, 2:17 doi:10.1186/1745-6150-2-17
Published: 28 June 2007Additional files
Additional file 1:
Mutations in neutrally evolved P450s. The text file gives the full sequence of the P450 R1-11 neutral evolution parent, as well as the individual mutations of all 34 neutrally evolved P450 variants. These data were originally reported in [16].
Format: TXT Size: 8KB Download file
Additional file 2:
Standard curves used to determine P450 activities. The PDF file shows all of the standard curves used to determine the P450 concentration and enzymatic activities. Points that were deemed to fall in the linear range, and so used to compute the standard curve slopes, are solid. Points that were deemed outside of the linear range are empty. Each curve shows the slopes computed for two different measurements of P450 concentration and enzymatic activity, and the average slope with standard error. These average slopes were used to compute the P450 activities.
Format: PDF Size: 223KB Download file
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Additional file 3:
Raw activity and sequence data. The text file gives the activities of the P450 variants on each of the six substrates, as measured in this study. It also lists the number of nonsynonymous mutations (M_NS) relative to the R1-11 neutral evolution parent sequence, as originally reported in [16]. Each row lists the data for a different P450 variant, and standard errors for the measured activities are shown in parentheses. Activities are the normalized standard curve slopes as described in the Methods.
Format: TXT Size: 3KB Download file
Additional file 4:
Variability of experimental measurements. The attached PDF file shows the results of repeated activity measurements performed on the R1-11 parent P450. The plots are scaled so that they can be directly compared to Figure 3 to see how much of observed differences among neutrally evolved P450 variants might be due to variability in experimental measurements. Such a comparison indicates that the experimental measurement variability is much less than the differences between P450s. The plots also show that most of the experimental variation is introduced during the activity measurements rather than during expression/lysis.
Format: PDF Size: 6KB Download file
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