Enzymatic activity versus structural dynamics: the case of acetylcholinesterase tetramer.

The function of many proteins, such as enzymes, is modulated by structural fluctuations. This is especially the case in gated diffusion-controlled reactions (where the rates of the initial diffusional encounter and of structural fluctuations determine the overall rate of the reaction) and in oligomeric proteins (where function often requires a coordinated movement of individual subunits). A classic example of a diffusion-controlled biological reaction catalyzed by an oligomeric enzyme is the hydrolysis of synaptic acetylcholine (ACh) by tetrameric acetylcholinesterase (AChEt). Despite decades of efforts, the extent to which enzymatic efficiency of AChEt (or any other enzyme) is modulated by flexibility is not fully determined. This article attempts to determine the correlation between the dynamics of AChEt and the rate of reaction between AChEt and ACh. We employed equilibrium and nonequilibrium electro-diffusion models to compute rate coefficients for an ensemble of structures generated by molecular dynamics simulation. We found that, for the static initial model, the average reaction rate per active site is approximately 22-30% slower in the tetramer than in the monomer. However, this effect of tetramerization is modulated by the intersubunit motions in the tetramer such that a complex interplay of steric and electrostatic effects either guides or blocks the substrate into or from each of the four active sites. As a result, the rate per active site calculated for some of the tetramer structures is only approximately 15% smaller than the rate in the monomer. We conclude that structural dynamics minimizes the adverse effect of tetramerization, allowing the enzyme to maintain similar enzymatic efficiency in different oligomerization states.

[1]  J. Sussman,et al.  The synaptic acetylcholinesterase tetramer assembles around a polyproline II helix , 2013 .

[2]  A. Goldman,et al.  Atomic structure of acetylcholinesterase from Torpedo californica: a prototypic acetylcholine-binding protein , 1991, Science.

[3]  B. Hasinoff Kinetics of acetylthiocholine binding to electric eel acetylcholinesterase in glycerol/water solvents of increased viscosity. Evidence for a diffusion-controlled reaction. , 1982, Biochimica et biophysica acta.

[4]  Nathan A. Baker,et al.  PDB2PQR: an automated pipeline for the setup of Poisson-Boltzmann electrostatics calculations , 2004, Nucleic Acids Res..

[5]  S. Moore,et al.  The peripheral anionic site of acetylcholinesterase: structure, functions and potential role in rational drug design. , 2006, Current pharmaceutical design.

[6]  Benzhuo Lu,et al.  Electrodiffusion: a continuum modeling framework for biomolecular systems with realistic spatiotemporal resolution. , 2007, The Journal of chemical physics.

[7]  P Taylor,et al.  Crystal Structure of Mouse Acetylcholinesterase , 1999, The Journal of Biological Chemistry.

[8]  J. Grassi,et al.  Conformational Flexibility of the Acetylcholinesterase Tetramer Suggested by X-ray Crystallography* , 1999, The Journal of Biological Chemistry.

[9]  Gerhard Klebe,et al.  PDB2PQR: expanding and upgrading automated preparation of biomolecular structures for molecular simulations , 2007, Nucleic Acids Res..

[10]  J Andrew McCammon,et al.  Feature-preserving adaptive mesh generation for molecular shape modeling and simulation. , 2008, Journal of molecular graphics & modelling.

[11]  Benzhuo Lu,et al.  Molecular surface-free continuum model for electrodiffusion processes. , 2008, Chemical physics letters.

[12]  Nathan A. Baker,et al.  Finite element analysis of the time-dependent Smoluchowski equation for acetylcholinesterase reaction rate calculations. , 2007, Biophysical journal.

[13]  J. Andrew McCammon,et al.  Diffusional dynamics of ligand-receptor association , 1986 .

[14]  J. Massoulie,et al.  The C-terminal peptides of acetylcholinesterase: cellular trafficking, oligomerization and functional anchoring. , 2005, Chemico-biological interactions.

[15]  Nathan A. Baker,et al.  Tetrameric mouse acetylcholinesterase: continuum diffusion rate calculations by solving the steady-state Smoluchowski equation using finite element methods. , 2005, Biophysical journal.

[16]  F. Rieger,et al.  Observation par microscopie électronique des formes allongées et globulaires de l'acétylcholinestérase de gymnote (Electrophorus electricus) , 1973 .

[17]  B. P. Doctor,et al.  Natural monomeric form of fetal bovine serum acetylcholinesterase lacks the C-terminal tetramerization domain. , 2003, Biochemistry.

[18]  Benzhuo Lu,et al.  Order N algorithm for computation of electrostatic interactions in biomolecular systems , 2006, Proceedings of the National Academy of Sciences.

[19]  P. Taylor,et al.  Crystal Structure of Mouse Acetylcholinesterase , 1999, The Journal of Biological Chemistry.

[20]  J. Mccammon,et al.  Gated binding of ligands to proteins , 1981, Nature.

[21]  J. Massoulie,et al.  Determinants of the t Peptide Involved in Folding, Degradation, and Secretion of Acetylcholinesterase* , 2005, Journal of Biological Chemistry.

[22]  D. Ermak,et al.  Brownian dynamics with hydrodynamic interactions , 1978 .

[23]  James Andrew McCammon,et al.  The Association of Tetrameric Acetylcholinesterase with ColQ Tail: A Block Normal Mode Analysis , 2005, PLoS Comput. Biol..

[24]  J. Mccammon,et al.  Dynamics of the Acetylcholinesterase Tetramer , 2007, Biophysical journal.

[25]  F. Rieger,et al.  Fine structure of electric eel acetylcholinesterase , 1975, Brain Research.

[26]  Nathan A. Baker,et al.  Finite element solution of the steady-state Smoluchowski equation for rate constant calculations. , 2004, Biophysical journal.

[27]  J. Mccammon,et al.  Stochastically gated diffusion‐influenced reactions , 1982 .

[28]  Benzhuo Lu,et al.  Continuum simulations of acetylcholine consumption by acetylcholinesterase: a Poisson-Nernst-Planck approach. , 2008, The journal of physical chemistry. B.

[29]  J. Sussman,et al.  Three-dimensional structures of acetylcholinesterase and of its complexes with anticholinesterase agents. , 1994, Biochemical Society transactions.