The University of Iowa

Daniel M. Quinn

Daniel M. Quinn
Professor
Phone: 
319-335-1335
Office: 
W333 CB
Office Hours: 
Mondays and Wednesdays 1:30p-3:00p or by appointment
Divisions: 
Biosketch: 
  • B.S., Quincy College (1972)
  • Ph.D., University of Kansas (1978)
  • Postdoctoral Research Associate, Indiana University (1978-80); NIH Postdoctoral Training Fellow, University of Cincinnati College of Medicine (1980-82)
Keywords: 

Molecular dynamics of acetylcholinesterase catalysis; mechanisms of enzymes of cardiovascular lipid metabolism; design of mechanism-based enzyme inhibitors; synthesis and evaluation of potential blood cholesterol lowering drugs.

Research Interests: 

It has long been appreciated that enzymes accelerate otherwise slow reactions by specific stabilization of the transition state. How this is achieved is still largely a matter of conjecture. My research group studies the catalytic mechanisms of three classes of biomedically relevant enzymes: cholinesterases, cholesterol esterases and b-lactamases. The overall aim of this research is to illuminate the intermolecular interactions that these enzyme utilize to effect molecular recognition of their substrates and concomitant transition state stabilization. A range of approaches is used to effect this aim, including measurement of structure-activity effects (e.g., kinetic isotope effects, free-energy relationships, site-specific mutagenesis), molecular modeling, and computational chemistry.

The physiological function of acetylcholinesterase (AChE) is the hydrolytic destruction of the neurotransmitter acetylcholine in the central and peripheral nervous systems, a task that the enzyme effects with very high efficiency. At low acetylcholine concentration, the reaction is rate-limited by the diffusional encounter of enzyme and substrate, while at high acetylcholine concentration, each AChE active site hydrolyzes 10,000 molecules of substrate per second! The surrogate chromogenic substrate acetylthiocholine is hydrolyzed by AChE as readily as acetylcholine, which provides a convenient analytical handle for characterizing catalysis. We are studying the diffusional encounter step by measuring ionic strength effects on rate constants for acetylthiocholine turnover and for binding and release of the tightly bound transition state analog inhibitor m-N,N,N-trimethylammonio-trifluoroacetophenone. At zero ionic strength, these rate constants are faster than the traditional speed limit of biological catalysis, which indicates that AChE uses an electrical field to accelerate binding of ligands to the active site. The effect of the electrical field is being further characterized by computing the lowest free energy pathway for cationic ligand diffusion through the active site gorge of the enzyme. The chemical steps that follow substrate binding to the active site are being characterized by measuring substrate and solvent isotope effects and by site-specific mutagenesis of active site residues. These experiments indicate that the enzyme stabilizes a quasi-tetrahedral transition state by a combination of proton transfer catalysis and electrostatic effects. Our investigations of AChE catalysis have been greatly informed by the availability of X-ray crystal structures for the free enzyme and for enzyme-ligand complexes (see Harel et al. in the accompanying references).

Cholesterol esterase (CEase) is a lipolytic enzyme that is involved in the digestion of dietary fats in the intestines and their subsequent absorption into the bloodstream. The physiological substrates of CEase are lipid esters, which include triacylglycerols and cholesteryl esters. This enzyme is thought to play a role in regulating the levels of low-density lipoproteins, the primary vehicles for carrying cholesterol through the circulatory system for delivery to peripheral tissues. We are investigating the ways in which CEase interacts with lipid ester substrates and with synthetic inhibitors. Because there is no X-ray structure for the enzyme, models for rat and human CEases have been constructed by homology modeling (see Feaster et al. in the accompanying references). These structural models aid interpretation of the effects of substrate and inhibitor structure on interaction with CEase. This combination of structure-activity experiments, inhibitor synthesis and computer modeling should promote the design of a CEase inhibitor that lowers serum cholesterol levels, and therefore is therapeutically useful in the treatment of heart disease.

Recent Publications: 
  • Tormos, J.R.; Wiley, K.L.; Schlom, P.S.; Wang, Y.; Fournier, D.; Masson, P.; Nachon, F.; Quinn, D.M.   Accumulation of Tetrahedral Intermediates in Cholinesterase Catalysis: a Secondary Isotope Effect Study. Journal of the American Chemical Society 2010, 132, 17751-17759.
  • Tormos, J.R.; Wiley, K.L.; Seravalli, J.; Nachon, F.; Masson, P.; Nicolet, Y.; Quinn, D.M.  The Reactant State for Substrate-Activated Turnover of Acetylthiocholine by Butyrylcholinesterase is a Tetrahedral Intermediate. Journal of the American Chemical Society 2005, 127,14538-14539.
  • Liang, Y.; Medhekar, R.; Brockman, H.L.; Quinn, D.M.; Hui, D.Y.   Importance of Arginines 63 and 423 in Modulating the Bile Salt-Dependent and Bile Salt-Independent Hydrolytic Activities of Rat Carboxyl Ester Lipase. Journal of Biological Chemistry 2000, 275, 24040-24046.
  • Malany, S.; Sawai, M.; Sikorski, R.S.; Seravalli, J.; Quinn, D.M.; Radić, Z.; Taylor, P.; Velan, B.; Kronman, C.; Shafferman, A.  Transition State Structure and Rate Determination for the Acylation Stage of Acetylcholinesterase Catalyzed Hydrolysis of (Acetylthio)choline. Journal of the American Chemical Society 2000, 122, 2981-2987.
  • Quinn, D.M.; Feaster, S.R.; Nair, H.K.; Baker, N.A.; Radić, Z.; Taylor, P.   Delineation and Decomposition of Energies Involved in Quaternary Ammonium  Binding in the Active Site of Acetylcholinesterase. Journal of the American Chemical Society 2000, 122, 2975-2980.
  • Feaster, S.R.; Quinn, D.M.; Barnett, B.L.  Molecular Modeling of the Structures of Human and Rat Pancreatic Cholesterol Esterases. Protein Science 1997, 6, 71-77.
  • Radić, Z.; Kirchhoff, P.; Quinn, D.M.; McCammon, J.A.; Taylor, P.   Electrostatic Influence on the Kinetics of Ligand Binding to Acetylcholinesterase:  Distinctions Between Active Center Ligands and Fasciculin. Journal of Biological Chemistry 1997, 272, 23265-23277.
  • Harel, M.; Quinn, D.M.; Nair, H.K.; Silman, I.; Sussman, J.L.  The X-ray Structure of a Transition State Analog Complex Reveals the Molecular Origins of the Catalytic Power and Substrate Specificity of Acetylcholinesterase. Journal of the American Chemical Society 2006, 118, 2340-2346.
  • Feaster, S.R.; Lee, K.; Baker, N.; Hui, D.Y.; Quinn, D.M.  Molecular Recognition by Cholesterol Esterase of Active Site Ligands:  Structure-Reactivity Effects for Inhibition by Aryl Carbamates and Subsequent Carbamylenzyme Turnover.  Biochemistry 1996, 35, 16723-16734.
  • Quinn, D.M.   Acetylcholinesterase:  Enzyme Structure, Reaction Dynamics, and Virtual Transition. States Chemical Reviews 1987, 87, 955-981.