Design of Protein–Peptide Interaction Modules for Assembling Supramolecular Structures in Vivo and in Vitro

• Published in: ACS chemical biology 2015-09, Vol.10 (9), p.2108-2115
• Main Authors: Speltz, Elizabeth B, Nathan, Aparna, Regan, Lynne
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Escherichia coli - chemistry; Escherichia coli - metabolism; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - metabolism; HEK293 Cells; HeLa Cells; Homeodomain Proteins - chemistry; Homeodomain Proteins - metabolism; HSP90 Heat-Shock Proteins - chemistry; HSP90 Heat-Shock Proteins - metabolism; Humans; Models, Molecular; Peptides - chemistry; Peptides - metabolism; Protein Interaction Domains and Motifs; Protein Interaction Maps; Proteins - chemistry; Proteins - metabolism; Tumor Suppressor Proteins - chemistry; Tumor Suppressor Proteins - metabolism
• ISSN: 1554-8929; 1554-8937
• DOI: 10.1021/acschembio.5b00415

Synthetic biology and protein origami both require protein building blocks that behave in a reliable, predictable fashion. In particular, we require protein interaction modules with known specificity and affinity. Here, we describe three designed TRAP (Tetratricopeptide Repeat Affinity Protein)–peptide interaction pairs that are functional in vivo. We show that each TRAP binds to its cognate peptide and exhibits low cross-reactivity with the peptides bound by the other TRAPs. In addition, we demonstrate that the TRAP–peptide interactions are functional in many cellular contexts. In extensions of these designs, we show that the binding affinity of a TRAP–peptide pair can be systematically varied. The TRAP–peptide pairs we present thus represent a powerful set of new building blocks that are suitable for a variety of applications.