Christian E. Schafmeister

Shape-Programmable Macromolecules

Proteins catalyze specific chemical reactions and carry out highly selective molecular recognition because they adopt well-defined three-dimensional structures and position chemically reactive functional groups in specific constellations. Proteins attain these well-defined structures through the complex process of protein folding. We seek to emulate these protein functions by constructing macromolecules that are easier to engineer by avoiding folding altogether. Toward that goal, we have developed an approach for the synthesis of macromolecules with programmable shapes. As described in this Account, we have constructed synthetic building blocks called bis-amino acids that we then couple through pairs of amide bonds to create water-soluble, spiroladder oligomers (bis-peptides) with well-defined three-dimensional structures. Bis-peptides use the conformational preferences of fused rings, stereochemistry, and strong covalent bonds to define their shape, unlike natural proteins and synthetic foldamers, which depend on noncovalent interactions and an unpredictable folding process to attain structure. Using these bis-amino acid monomers, we have built and characterized a number of bis-peptide nanostructures. We also constructed a molecular actuator that undergoes a large change in conformation under the control of metal exchange; the first application of bis-peptides. We are currently developing further approaches to functionalize bis-peptides as scaffolds to present well-defined constellations of functional groups. Such macromolecules could facilitate multifunctional catalysis and molecular recognition and lead to nanoscale molecular devices.

Hydrophobic Substituent Effects on Proline Catalysis of Aldol Reactions in Water

Derivatives of 4-hydroxyproline with a series of hydrophobic groups in well-defined orientations have been tested as catalysts for the aldol reactions. All of the modified proline catalysts carry out the intermolecular aldol reaction in water and provide high diastereoselectivity and enantioselectivity. Modified prolines with aromatic groups syn to the carboxylic acid are better catalysts than those with small hydrophobic groups (1a is 43.5 times faster than 1f). Quantum mechanical calculations provide transition structures, TS-1a(water) and TS-1f(water), that support the hypothesis that a stabilizing hydrophobic interaction occurs with 1a.

Spiroligozymes for transesterifications: design and relationship of structure to activity.

Transesterification catalysts based on stereochemically defined, modular, functionalized ladder-molecules (named spiroligozymes) were designed, using the inside-out design strategy, and mutated synthetically to improve catalysis. A series of stereochemically and regiochemically diverse bifunctional spiroligozymes were first synthesized to identify the best arrangement of a pyridine as a general base catalyst and an alcohol nucleophile to accelerate attack on vinyl trifluoroacetate as an electrophile. The best bifunctional spiroligozyme reacted with vinyl trifluoroacetate to form an acyl-spiroligozyme conjugate 2.7 × 10(3)-fold faster than the background reaction with a benzyl alcohol. Two trifunctional spiroligozymes were then synthesized that combined a urea with the pyridine and alcohol to act as an oxyanion hole and activate the bound acyl-spiroligozyme intermediate to enable acyl-transfer to methanol. The best trifunctional spiroligozyme carries out multiple turnovers and acts as a transesterification catalyst with k(1)/k(uncat) of 2.2 × 10(3) and k(2)/k(uncat) of 1.3 × 10(2). Quantum mechanical calculations identified the four transition states of the catalytic cycle and provided a detailed view of every stage of the transesterification reaction.

Synthesis of hexa- and pentasubstituted diketopiperazines from sterically hindered amino acids.

Steric hindrance assists in the formation of hindered diketopiperazines using acyl-transfer coupling. In acyl-transfer coupling, the carboxylate of an unprotected N-alkylamino acid attacks an active ester to form a transient anhydride that undergoes an O,N acyl transfer to form a tertiary amide. If the active ester is part of an N-alkylamino acid it will form a diketopiperazine. It is demonstrated here that acyl-transfer coupling can assemble highly functionalized bis-peptides bearing a functional group on every monomer.

Determination of Internal Energy Distributions of Laser Electrospray Mass Spectrometry using Thermometer Ions and Other Biomolecules

AbstractThe internal energy distributions for dried and liquid samples that were vaporized with femtosecond duration laser pulses centered at 800xa0nm and postionized by electrospray ionization-mass spectrometry (LEMS) were measured and compared with conventional electrospray ionization mass spectrometry (ESI-MS). The internal energies of the mass spectral techniques were determined by plotting the ratio of the intact parent molecular features to all integrated ion intensities of the fragments as a function of collisional energy using benzylpyridinium salts and peptides. Measurements of dried p-substituted benzylpyridinium salts using LEMS resulted in a greater extent of fragmentation in addition to the benzyl cation. The mean relative internal energies, were determined to be 1.62u2009±u20090.06, 2.0u2009±u20090.5, and 1.6u2009±u20090.3xa0eV for ESI-MS, dried LEMS, and liquid LEMS studies, respectively. Two-photon resonances with the laser pulses likely caused lower survival yields in LEMS analyses of dried samples but not liquid samples. In studies with larger biomolecules, LEMS analyses of dried samples from glass showed a decrease in survival yield compared with conventional ESI-MS for leucine enkephalin and bradykinin of ~15% and 11%, respectively. The survival yields for liquid LEMS analyses were comparable to or better than ESI-MS for benzylpyridinium salts and large biomolecules.n Figureᅟ

Synthesis of a Carboxylate Functionalized Bis-Amino Acid Monomer

The synthesis of the first functionalized bis-amino acid monomer proAc(2S3S4R) 1 that carries an acetyl side chain is presented. This monomer was incorporated into oligomer 3 and the solution phase structure was determined by using two-dimensional nuclear magnetic resonance. The solution structure confirmed the intended connectivity and stereochemistry of the oligomer. This first functionalized bis-amino acid represents a milestone toward functionalized bis-peptide nanostructures for catalytic, molecular recognition, and nanotechnology applications.

Experimental evidence for water mediated electron transfer through bis-amino acid donor-bridge-acceptor oligomers.

This work compares the photoinduced unimolecular electron transfer rate constants for two different solute molecules (D-SSS-A and D-SRR-A) in water and DMSO solvents. The D-SSS-A solute has a cleft between the electron donor and acceptor units, which is able to contain a water molecule but is too small for DMSO. The rate constant for D-SSS-A in water is significantly higher than that for D-SRR-A, which lacks a cleft, and significantly higher for either solute in DMSO. The enhancement of the rate constant is explained by an electron tunneling pathway that involves water molecule(s).

Acceleration of an aromatic Claisen rearrangement via a designed spiroligozyme catalyst that mimics the ketosteroid isomerase catalytic dyad.

A series of hydrogen-bonding catalysts have been designed for the aromatic Claisen rearrangement of a 1,1-dimethylallyl coumarin. These catalysts were designed as mimics of the two-point hydrogen-bonding interaction present in ketosteroid isomerase that has been proposed to stabilize a developing negative charge on the ether oxygen in the migration of the double bond.1 Two hydrogen bond donating groups, a phenol alcohol and a carboxylic acid, were grafted onto a conformationally restrained spirocyclic scaffold, and together they enhance the rate of the Claisen rearrangement by a factor of 58 over the background reaction. Theoretical calculations correctly predict the most active catalyst and suggest that both preorganization and favorable interactions with the transition state of the reaction are responsible for the observed rate enhancement.

Observation of Contraction and Expansion in a Bis(peptide)‐Based Mechanical Molecular Actuator

A novel, bis(peptide) based molecular actuator (1) has been synthesized. It is demonstrated to undergo contraction and expansion controlled by the addition and removal of Cu2+; this is demonstrated by the direct observation of a change in hydrodynamic properties by using sedimentation analysis and size exclusion chromatography. The molecule undergoes a large change in sedimentation coefficient, axial ratio, and size exclusion chromatography elution time when it binds copper. The demonstration of a controlled change in the mechanical properties of 1 make it a good starting point for the development of molecular devices that will harness changes in molecular shape and size to create molecular devices such as sensors or valves.

Synthesis of Spiroligomer-Containing Macrocycles.

We demonstrate the synthesis and characterization of the solution conformations of a collection of functionalized spiroligomer-based macrocycles. These macrocycles contain 14 independently controllable stereocenters and four independently controllable functional groups on a highly preorganized scaffold. These molecules are being developed to display complex, preorganized surfaces for binding proteins and to create enzyme-like active sites. In this work, we demonstrate the convergent synthetic approach to this new class of macrocycles and demonstrate that the conformational properties of these molecules can be changed by altering the configuration stereocenters within the backbone.