Free energy calculations on binding and catalysis by α-lytic protease: The role of substrate size in the P1 pocket

✍ Scribed by James W. Caldwell; David A. Agard; Dr. Peter A. Kollman

• Book ID: 105358564
• Publisher: John Wiley and Sons
• Year: 1991
• Tongue: English
• Weight: 594 KB
• Volume: 10
• Category: Article
• ISSN: 0887-3585
• DOI: 10.1002/prot.340100207

✦ Synopsis

We present free energy calculations using molecular dynamics on different substrates of alpha-lytic protease in the gas phase, in solution, while forming a noncovalent Michaelis complex with the enzyme, and in a tetrahedral structure representing a transition state/intermediate for acylation by the enzyme. Various P1 substrates were studied, with P1 = Gly, Ala, Val, and Leu. In qualitative agreement with experiment, the enzyme was calculated to bind and catalyze most effectively substrates with P1 = Ala over those with P1 = Gly, Val or Leu. Also, the calculated relative solvation free energies of Gly----Ala and Ala----Val were in qualitative agreement with experimental values in corresponding model systems. However, the level of quantitative agreement with experiment achieved in our earlier study of relative binding and catalysis of native subtilisin and an Asn-155----Ala mutant was not achieved. We surmise that this is due to the greater difficulty in quantitatively simulating effects that are predominantly van der Waals and hydrophobic compared to those that are hydrogen bonding/electrostatic.