George Whitesides and his lab have another paper out on the details of how ligands bind to proteins. They're still using the favorite model enzyme of all time (carbonic anhydrase), the fruit fly and nematode of the protein world. Last time around, using a series of ligands and their analogs with an extra phenyl in their structure. The benzo-ligands had increased affinity, and this seemed to be mostly an enthalpic effect. After a good deal of calorimetry, etc., they concluded that the balancing act between enthalpy and entropy they saw over the group was different for ligand binding than it was for logP partitioning, and that means that it doesn't really match up with the accepted definition of a "hydrophobic effect".
In this study, they're looking at fluorinated analogs of the same compounds to see what that might do to the binding process. That makes the whole thing interesting for a medicinal chemist, because we make an awful lot of fluorinated analogs. You can start some interesting discussions about whether these are more hydrophobic than their non-F analogs, though, and this paper lands right in the middle of that issue.
The first result was that the fluorinated analogs bound to the enzyme (in their X-ray structures) with almost identical geometry. That makes the rest of the discussion easier to draw conclusions from (and more relevant). It's worth remembering, though, that very small changes can still add up. There was a bit of a shift in the binding pocket, actually, which they attribute to an unfavorable interaction between the fluorines and the carbonyl of a threonine residue. But the carbonic anhydrase pocket is pretty accomodating - the overall affinity of the compounds did not really change. That led to this conclusion:
Values of DG8bind, combined with an overall conserved binding geometry of each set of benzo- and fluorobenzo-extended ligands suggest that binding depends on a fine balance of interactions between HCA, the ligand, and molecules of water filling the pocket and surrounding the ligand, and that a simple analysis of interactions between the protein and ligand (Figure1E) is insufficient to understand (or more importantly, predict) the free energy of binding.
But although the overall free energy didn't change, the enthalpic and entropic components did (but arrived at the same place, another example to add to the long list of systems that do this). The differences seem to be in the Coulombic interaction with the binding pocket (worse enthalpy term - is that what shifted the structure over a bit in the X-ray?) and changes in energy of solvation as the ligand binds (better entropy term). Matched pairs of compounds didn't really show a difference in how many waters they displaced from the binding site.
So the take-home is that the hydrophobic effect is not all about releasing waters from protein binding surfaces, as has been proposed by some. It's a mixture of stuff, and especially depends on the structure of the water in the binding pocket and around the ligands, and the changes in these as the compounds leave bulk solvent and find their way into the binding site.
That makes things tricky for many compounds. Hydrophobic effects seem to be a big part of the binding energy of a lot of drug molecules (despite various efforts to cut back on this), and these Whitesides studies would say that modeling and predicting these energetic changes are going to be hard. Computationally, we'd have an easier time figuring out direct interactions between the protein and the ligand, the way we do with enthalpic interactions like hydrogen bonds. Keeping track of all those water molecules is more painful - but necessary.