Evolution favors protein mutational robustness in sufficiently large populations

Additional file 1:

Sequence of the parent P450 used start neutral evolution. FASTA file with sequence of the R1-11 P450 BME used as the neutral evolution parent. This sequence was isolated after the equilibration evolution.

Format: FASTA Size: 2KB Download file

Additional file 2:

Information about sequences from neutral evolution experiments. The entries give the name of the mutant, the number of nonsynonymous and nucleotide mutations relative to the R1-11 parent, the [urea]₅₀ value if measured, the T₅₀ value if measured, the percent of the parental expression level if measured, and then a list of all of the mutations. Amino acid mutations are numbered in the standard P450 numbering scheme. The names of the mutants indicate their origin. Names beginning with "P-G3" are randomly chosen functional mutants from generation 3 of the polymorphic population, etc. Names of the form "P1," "P2,", etc. are the 22 functional mutants that were randomly chosen from the final (generation 15) polymorphic population. Numbers P5 and P12 are missing because two of the original 24 randomly selected polymorphic population members were randomly chosen to be discarded after it was discovered that two of the 24 monomorphic replicates were contaminated. Names beginning with "U1" indicate that sequences are from the first unselected replicate, etc. Names beginning "M1" indicate sequences are from the first monomorphic replicate, etc. Replicates "M9" and "M10" were discarded due to contamination during the experiment. For each replicate, we sequenced each new functional mutant. The last functional mutant after 25 generations represents the final sequence for that replicate, and is given an abbreviated name without the generation suffix.

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Additional file 3:

Thermostability measurements. Raw data from the T₅₀ thermostability measurements.

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Additional file 4:

Urea stability measurements. Raw data from the [urea]₅₀ thermostability measurements.

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Additional file 5:

Correlation of thermal and urea stabilities. The T₅₀ and [urea]₅₀ values are highly correlated.

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Additional file 6:

Sequence of initial P450 used to start equilibration evolution. FASTA file with sequence of the 21B3 P450 BM3 heme domain described in [23]. This P450 was used as the initial parent to start the equilibration evolution.

Format: FASTA Size: 2KB Download file

Additional file 7:

Mutations accumulated during equilibration evolution. The file lists the mutations in the 46 P450 variants selected at the end of the equilibration evolution. Each line gives the name of the variant, with the prefix indicating whether it came from the R1 or R2 population. The next entries give the number of nucleotide and nonsynonymous mutations. All of the individual mutations relative to 21B3 are then listed. Amino acid mutations are numbered in the standard P450 numbering scheme, with the threonine after the N-terminal methionine given the number one.

Format: TXT Size: 20KB Download file