title: Computational design of receptors for an organophosphate surrogate of the nerve agent soman.
abstract: We report the computational design of soluble protein receptors for pinacolyl methyl phosphonic acid (PMPA), the predominant hydrolytic product of the nerve agent soman. Using recently developed computational protein design techniques, the ligand-binding pockets of two periplasmic binding proteins, glucose-binding protein and ribose-binding protein, were converted to bind PMPA instead of their cognate sugars. The designs introduce 9-12 mutations in the parent proteins. Twelve of 20 designs tested exhibited PMPA-dependent changes in emission intensity of a fluorescent reporter with affinities between 45 nM and 10 microM. The contributions to ligand binding by individual residues were determined in two designs by alanine-scanning mutagenesis, and are consistent with the molecular models. These results demonstrate that designed receptors with radically altered binding specificities and affinities that rival or exceed those of the parent proteins can be successfully predicted. The designs vary in parent scaffold, sequence diversity, and orientation of docked ligand, suggesting that the number of possible solutions to the design problem is large and degenerate. This observation has implications for the genesis of biological function by random processes. The designed receptors reported here may have utility in the development of fluorescent biosensors for monitoring nerve agents.
authors.affiliation: Departments of Biochemistry and Pharmacology and Molecular Cancer Biology, Box 3711, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Malin
authors.initials: M
authors.lastname: Allert
authors.forename: Shahir S
authors.initials: SS
authors.lastname: Rizk
authors.forename: Loren L
authors.initials: LL
authors.lastname: Looger
authors.forename: Homme W
authors.initials: HW
authors.lastname: Hellinga
mesh_terms: D001665:Binding Sites; D015374:Biosensing Techniques; D003198:Computer Simulation; D015195:Drug Design; D008024:Ligands; D008958:Models, Molecular; D016296:Mutagenesis / Q000235:genetics; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012999:Soman / Q000031:analogs & derivatives* / Q000378:metabolism*; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D008024:Ligands; D011956:Receptors, Cell Surface; C046573:pinacolyl methylphosphonic acid; D012999:Soman