title: Synthesis and folding of a mirror-image enzyme reveals ambidextrous chaperone activity.
abstract: Mirror-image proteins (composed of D-amino acids) are promising therapeutic agents and drug discovery tools, but as synthesis of larger D-proteins becomes feasible, a major anticipated challenge is the folding of these proteins into their active conformations. In vivo, many large and/or complex proteins require chaperones like GroEL/ES to prevent misfolding and produce functional protein. The ability of chaperones to fold D-proteins is unknown. Here we examine the ability of GroEL/ES to fold a synthetic d-protein. We report the total chemical synthesis of a 312-residue GroEL/ES-dependent protein, DapA, in both L- and D-chiralities, the longest fully synthetic proteins yet reported. Impressively, GroEL/ES folds both L- and D-DapA. This work extends the limits of chemical protein synthesis, reveals ambidextrous GroEL/ES folding activity, and provides a valuable tool to fold d-proteins for drug development and mirror-image synthetic biology applications.
authors: Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84112-5650.
authors: Matthew T
authors: MT
authors: Weinstock
authors: Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84112-5650.
authors: Michael T
authors: MT
authors: Jacobsen
authors: Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84112-5650 kay@biochem.utah.edu.
authors: Michael S
authors: MS
authors: Kay
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D055592:Biophysical Phenomena; D018835:Chaperonin 10 / Q000378:metabolism; D018834:Chaperonin 60 / Q000378:metabolism; D004798:Enzymes / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006836:Hydro-Lyases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000378:metabolism*; D008969:Molecular Sequence Data; D010848:Picolinic Acids / Q000378:metabolism; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D013237:Stereoisomerism
chemical_list: C550634:4-hydroxy-2,3,4,5-tetrahydro-2-dipicolinic acid; D000596:Amino Acids; D018835:Chaperonin 10; D018834:Chaperonin 60; D004798:Enzymes; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D010848:Picolinic Acids; D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase
keywords: peptide synthesis; protein folding