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. 2021 Jun 10:8:654706.
doi: 10.3389/fmolb.2021.654706. eCollection 2021.

Thermodynamic Analysis for Binding of 4- O-β-tri- N-acetylchitotriosyl Moranoline, a Transition State Analogue Inhibitor for Hen Egg White Lysozyme

Makoto Ogata  1 , Tamo Fukamizo  2 , Takayuki Ohnuma  2   3
Affiliations

Thermodynamic Analysis for Binding of 4- O-β-tri- N-acetylchitotriosyl Moranoline, a Transition State Analogue Inhibitor for Hen Egg White Lysozyme

Makoto Ogata et al. Front Mol Biosci. .

Abstract

4-O-β-tri-N-acetylchitotriosyl moranoline (GN3M) is a transition-state analogue for hen egg white lysozyme (HEWL) and identified as the most potent inhibitor till date. Isothermal titration calorimetry experiments provided the thermodynamic parameters for binding of GN3M to HEWL and revealed that the binding is driven by a favorable enthalpy change (ΔH° = -11.0 kcal/mol) with an entropic penalty (-TΔS° = 2.6 kcal/mol), resulting in a free energy change (ΔG°) of -8.4 kcal/mol [Ogata et al. (2013) 288, 6,072-6,082]. Dissection of the entropic term showed that a favorable solvation entropy change (-TΔS solv° = -9.2 kcal/mol) is its sole contributor. The change in heat capacity (ΔC p°) for the binding of GN3M was determined to be -120.2 cal/K·mol. These results indicate that the bound water molecules play a crucial role in the tight interaction between GN3M and HEWL.

Keywords: 4-O-β-tri-N-acetylchitotriosyl moranoline; binding; inhibitor; lysozyme (HEWL); thermodynamics.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Chemical structure of GN3M and (GlcNAc)3.
FIGURE 2
FIGURE 2
ITC thermogram (upper) and theoretical fit to the experimental data (lower) for binding of GN3M at 20°C. Inset shows the bar diagram of the thermodynamic parameters (A). Temperature dependence of GN3M binding to HEWL; the plot of ΔH° versus temperature yielded a change in the heat capacity (ΔC p°) based on the slope of the line. The ΔC p° value was calculated to be −120.2 cal/K·mol (B).
FIGURE 3
FIGURE 3
Crystal structures of (GlcNAc)3-liganded and GN3M-liganded HEWL (A) Stereo view of superimposed structures of (GlcNAc)3-liganded HEWL (green; PDB code 1lzb) and GN3M-liganded HEWL (cyan; PDB code 4hp0) complexes. The catalytic residues Glu35 and Asp52 of HEWL are indicated as sticks (GlcNAc)3 and GN3M are shown as orange and yellow sticks, respectively. (B) The binding modes of (GlcNAc)3 and GN3M to HEWL. Amino acid residues involved in the binding of ligands (GlcNAc)3 and GN3M are also indicated as sticks. The numbers, −4 to −1, indicate the subsite positions. Dashed lines indicate the possible hydrogen bonds. Red spheres represent oxygen atoms of water molecules.
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