Anilinonaphthalenesulfonate (ANS) and tryptophan compete for binding to the trp repressor protein; thus, the fluorescence decrease associated with ANS dissociation can be used as a fluorometric marker for tryptophan binding to the protein. Using this approach, the tryptophan equilibrium dissociation constant was measured at 25 degrees C to be 3.7 (+/- 1.2) X 10(-5) M, a value which compares fav...

The protonation constant of hexamethylenetetramine (urotropine) was determined by a potentiometric and a spectrophotometric method. The calculations gave log K(HL) (concentration constants): 4.89 at mu = 0.1 and 5.05 at mu = 0.5. The temperature was 25 degrees and potassium chloride was used to adjust the ionic strength.

The photophysics and spectroscopy of tryptophylglycine, tryptophylalanine, glycyltryptophan, alanyltryptophan, glycyltryptophylglycine, and tryptophan itself have been investigated by using steady-state and subnanosecond spectroscopy. For tryptophan and the peptides where the tryptophyl residue is N-terminal we demonstrate the involvement of the state of protonation on both the excited-state dy...

A new method is proposed for constant pH molecular dynamics (MD), employing generalized Born (GB) electrostatics. Protonation states are modeled with different charge sets, and titrating residues sample a Boltzmann distribution of protonation states as the simulation progresses, using Monte Carlo sampling based on GB-derived energies. The method is applied to four different crystal structures o...

Redox-active tryptophans are important in biological electron transfer and redox biochemistry. Proteins can tune the electron transfer kinetics and redox potentials of tryptophan via control of the protonation state and the hydrogen-bond strength. We examine the local environment of two neutral tryptophan radicals (Trp108 on the solvent-exposed surface and Trp48 buried in the hydrophobic core) ...

Compstatin is a 13-residue cyclic peptide that inhibits complement activation by binding to complement component, C3. Although the activity of compstatin has been improved severalfold using combinatorial and rational design approaches, the molecular basis for its interaction with C3 is not yet fully understood. In the present study, isothermal titration calorimetry was employed to dissect the m...

We have studied the apo (Fe(3+) free) form of periplasmic ferric binding protein (FbpA) under different conditions and we have monitored the changes in the binding and release dynamics of H2PO4(-) that acts as a synergistic anion in the presence of Fe(3+). Our simulations predict a dissociation constant in agreement with the experimentally measured value under the same ionic pH conditions. We a...

pH is an important parameter in condensed-phase systems, because it determines the protonation state of titratable groups and thus influences the structure, dynamics, and function of molecules in solution. In most force field simulation protocols, however, the protonation state of a system (rather than its pH) is kept fixed and cannot adapt to changes of the local environment. Here, we present ...

To understand ion permeation, one must assign correct ionization states to titratable amino acid residues in protein channels. We report on the effects of physical and methodological assumptions in calculating the protonation states at neutral bulk pH of titratable residues lining the lumen of the native Escherichia coli OmpF channel, and five mutants. We systematically considered a wide range ...