Identification of N-terminal helix capping boxes by means of <Superscript>13 </Superscript>C chemical shifts

We have examined the 13C~ and 13C[3 chemical shifts of a number of proteins and found that their values at the N-terminal end of a helix provide a good predictor for the presence of a capping box. A capping box consists of a hydrogen-bonded cycle of four amino acids in which the side chain of the N-cap residue forms a hydrogen bond with the backbone amide of the N3 residue, whose side chain in turn may accept a hydrogen bond from the amide of the N-cap residue. The N-cap residue exhibits characteristic values for its backbone torsion angles, with r and ~ clustering around 94 + 15 ~ and 167 + 5 ~ respectively. This is manifested by a 1-2 ppm upfield shift of the 13C~ resonance and a 1 4 ppm downfield shift of the ~3C~ resonance, relative to their random coil values, and is mainly associated with the unusually large value of~. The residues following the N-cap residue exhibit downfield shifts of 1-3 ppm for the 13C~ resonances and small upfield shifts for the 13C13 ones, typical of an a-helix.