title: Macromolecular Crystallography for Synthetic Abiological Molecules: Combining xMDFF and PHENIX for Structure Determination of Cyanostar Macrocycles.
abstract: Crystal structure determination has long provided insight into structure and bonding of small molecules. When those same small molecules are designed to come together in multimolecular assemblies, such as in coordination cages, supramolecular architectures and organic-based frameworks, their crystallographic characteristics closely resemble biological macromolecules. This resemblance suggests that biomacromolecular refinement approaches be used for structure determination of abiological molecular complexes that arise in an aggregate state. Following this suggestion we investigated the crystal structure of a pentagonal macrocycle, cyanostar, by means of biological structure analysis methods and compared results to traditional small molecule methods. Cyanostar presents difficulties seen in supramolecular crystallography including whole molecule disorder and highly flexible solvent molecules sitting in macrocyclic and intermolecule void spaces. We used the force-field assisted refinement method, molecular dynamics flexible fitting algorithm for X-ray crystallography (xMDFF), along with tools from the macromolecular structure determination suite PHENIX. We found that a standard implementation of PHENIX, namely one without xMDFF, either fails to produce a solution by molecular replacement alone or produces an inaccurate structure when using generic geometry restraints, even at a very high diffraction data resolution of 0.84 Å. The problems disappear when taking advantage of xMDFF, which applies an optimized force field to realign molecular models during phasing by providing accurate restraints. The structure determination for this model system shows excellent agreement with the small-molecule methods. Therefore, the joint xMDFF-PHENIX refinement protocol provides a new strategy that uses macromolecule methods for structure determination of small molecules and their assemblies.
authors.affiliation: †Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, 405 North Mathews Avenue, Urbana, Illinois 61801, United States.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Singharoy
authors.affiliation: ‡Molecular and Cellular Biochemistry Department, Indiana University, 212 South Hawthorne Drive, Bloomington, Indiana 47405, United States.
authors.forename: Balasubramanian
authors.initials: B
authors.lastname: Venkatakrishnan
authors.affiliation: §Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405, United States.
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.affiliation: †Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, 405 North Mathews Avenue, Urbana, Illinois 61801, United States.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Mayne
authors.affiliation: §Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405, United States.
authors.forename: Semin
authors.initials: S
authors.lastname: Lee
authors.affiliation: §Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405, United States.
authors.forename: Chun-Hsing
authors.initials: CH
authors.lastname: Chen
authors.affiliation: ‡Molecular and Cellular Biochemistry Department, Indiana University, 212 South Hawthorne Drive, Bloomington, Indiana 47405, United States.
authors.forename: Adam
authors.initials: A
authors.lastname: Zlotnick
authors.affiliation: †Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, 405 North Mathews Avenue, Urbana, Illinois 61801, United States.|∥Department of Physics, University of Illinois at Urbana-Champaign, 1110 West Green Street, Urbana, Illinois 61801, United States.
authors.forename: Klaus
authors.initials: K
authors.lastname: Schulten
authors.affiliation: §Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405, United States.
authors.forename: Amar H
authors.initials: AH
authors.lastname: Flood
mesh_terms: D018360:Crystallography, X-Ray; D047028:Macrocyclic Compounds / Q000737:chemistry*; D008958:Models, Molecular; D008968:Molecular Conformation
chemical_list: D047028:Macrocyclic Compounds