An Amino Acid Packing Code for α-Helical Structure and Protein Design

Abstract

This work demonstrates that all packing in α-helices can be simplified to repetitive patterns of a single motif: the knob–socket. Using the precision of Voronoi Polyhedra/Delauney Tessellations to identify contacts, the knob–socket is a four-residue tetrahedral motif: a knob residue on one α-helix packs into the three-residue socket on another α-helix. The principle of the knob–socket model relates the packing between levels of protein structure: the intra-helical packing arrangements within secondary structure that permit inter-helix tertiary packing interactions. Within an α-helix, the three-residue sockets arrange residues into a uniform packing lattice. Inter-helix packing results from a definable pattern of interdigitated knob–socket motifs between two α-helices. Furthermore, the knob–socket model classifies three types of sockets: (1) free, favoring only intra-helical packing; (2) filled, favoring inter-helical interactions; and (3) non, disfavoring α-helical structure. The amino acid propensities in these three socket classes essentially represent an amino acid code for structure in α-helical packing. Using this code, we used a novel yet straightforward approach for the design of α-helical structure to validate the knob–socket model. Unique sequences for three peptides were created to produce a predicted amount of α-helical structure: mostly helical, some helical, and no helix. These three peptides were synthesized, and helical content was assessed using CD spectroscopy. The measured α-helicity of each peptide was consistent with the expected predictions. These results and analysis demonstrate that the knob–socket motif functions as the basic unit of packing and presents an intuitive tool to decipher the rules governing packing in protein structure.