TY - JOUR
T1 - Evidence for strained interactions between side-chains and the polypeptide backbone
AU - Stites, Wesley E.
AU - Meeker, Alan K.
AU - Shortle, David
N1 - Funding Information:
We are indebted to Neil Clarke and Apo Gittis for their thoughtful advice and help with calculations. We thank Lothar Sch~fer and Julian Pranata for critical reading of the manuscript. We thank Janet Thornton and Malcolm MacArthur for unpublished data on ~b,~b angle distribution in the protein databank. This work was supported by NIH grant GM34171 to D. Shortle. W.E.S. was supported by a NIH postdoctoral fellowship.
PY - 1994/1/7
Y1 - 1994/1/7
N2 - In most proteins, a small but significant fraction of residues adopt φ,ψ angles that generate unfavorable steric interactions between side-chain atoms and the peptide backbone. For the small protein staphylococcal nuclease, the X-ray structure reveals that 18 of 133 residues occupy unusual and, presumably, energetically unfavorable backbone conformations. To quantify the amount of strain energy generated by these local interactions, we have analyzed the changes in stability that accompany replacement of the wild-type side-chain with glycine, a residue that can access a much larger set of φ,ψ angles without energy penalty. To correct for the many other sources of stability loss that might accompany this mutation, the glycine mutant was compared to an alanine mutant at the same position and the resulting free energy difference ΔΔGG→A was then compared to the average ΔΔGG→A at all other, unstrained positions in the nuclease occupied by similar amino acid types. In addition, potential steric clashes were introduced by substituting alanine at each of six positions occupied in the wild-type by glycine with φ,ψ angles that are unfavorable for all other residue types. The data suggest that residues with φ,ψ angles outside the preferred α-helical and β-sheet regions represent sites of local strain energy that lower the stability of the native state by 1 to 2 kcal/mol and, in some cases, as much as 3 to 4 kcal/mol. Given that 10 to 20% of residues in globular proteins adopt φ,ψ angles outside the preferred α-helical and β-sheet regions, this implies that there is on the order of 20 kcal/mol of strain energy in a protein of 100 residues that may be relieved by appropriate mutations.
AB - In most proteins, a small but significant fraction of residues adopt φ,ψ angles that generate unfavorable steric interactions between side-chain atoms and the peptide backbone. For the small protein staphylococcal nuclease, the X-ray structure reveals that 18 of 133 residues occupy unusual and, presumably, energetically unfavorable backbone conformations. To quantify the amount of strain energy generated by these local interactions, we have analyzed the changes in stability that accompany replacement of the wild-type side-chain with glycine, a residue that can access a much larger set of φ,ψ angles without energy penalty. To correct for the many other sources of stability loss that might accompany this mutation, the glycine mutant was compared to an alanine mutant at the same position and the resulting free energy difference ΔΔGG→A was then compared to the average ΔΔGG→A at all other, unstrained positions in the nuclease occupied by similar amino acid types. In addition, potential steric clashes were introduced by substituting alanine at each of six positions occupied in the wild-type by glycine with φ,ψ angles that are unfavorable for all other residue types. The data suggest that residues with φ,ψ angles outside the preferred α-helical and β-sheet regions represent sites of local strain energy that lower the stability of the native state by 1 to 2 kcal/mol and, in some cases, as much as 3 to 4 kcal/mol. Given that 10 to 20% of residues in globular proteins adopt φ,ψ angles outside the preferred α-helical and β-sheet regions, this implies that there is on the order of 20 kcal/mol of strain energy in a protein of 100 residues that may be relieved by appropriate mutations.
KW - Ramachandran plot
KW - mutagenesis
KW - protein folding
KW - staphylococcal nuclease
KW - steric interactions
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U2 - 10.1016/S0022-2836(05)80008-7
DO - 10.1016/S0022-2836(05)80008-7
M3 - Article
C2 - 8289248
AN - SCOPUS:0028014680
SN - 0022-2836
VL - 235
SP - 27
EP - 32
JO - Journal of molecular biology
JF - Journal of molecular biology
IS - 1
ER -