Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Protein-ligand interaction investigated by HSQC titration study

  • Lee, Joon-Hwa (Department of Chemistry and RINS, Gyeongsang National University)
  • Received : 2018.11.16
  • Accepted : 2018.12.15
  • Published : 2018.12.20
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Abstract

Chemical shift perturbation (CSP) is a simple NMR technique for studying binding of a protein to various ligands. CSP is the only technique that can directly provide both a value for the dissociation constant and a binding site from the same set of measurements. To accurately analyze the CSP data, the exact binding mode such as multiple binding, should be carefully considered. In this review, we analyzed systematically the CSP data with multiple modes. This analysis might provide insight into the mechanism on how proteins selectively recognize their target ligands to achieve the biological function.

Keywords

JGGMB2_2018_v22n4_125_f0001.png 이미지

Figure 1. One-site binding model. (A) The simulated titration curves for P with L as a function of [L]t/[P]t ratio using Eq. 14. The colors used to illustrate the Kd are: red, 0.1 μM; blue, 1 μM; green, 10 μM; orange, 100 μM. The initial value of [P]t is 0.1 mM. (B) The calculated 1H/15N-HSQC cross-peaks of P during titration with L using Kd of (a) 0.1 μM, (b) 1 μM, (c) 10 μM, or (d) 100 μM. The cross-peak color changes gradually from blue to purple according to the [L]t/[P]t ratio. The black cross-peak indicates the bound state (PL).

JGGMB2_2018_v22n4_125_f0002.png 이미지

Figure 2. Two-site binding model: one protein and two ligands. (A) The calculated 1H/15N-HSQC cross-peaks of P during titration with L using Kd,2 of 1 μM and Kd,1 of (a) 0.01 μM, (b) 0.1 μM, (c) 1 μM, or (d) 10 μM. The cross-peak color changes gradually from blue to purple according to the [L]t/[P]t ratio. The black cross-peak indicates the two bound states (PL and PL2). (B) The simulated titration curves for P (a: peak A; b: peak B in Figure 2A) with L as a function of [L]t/[P]t ratio using Eq. 29 (Kd,2 = 1 μM). The colors used to illustrate the Kd,1 are: red, 0.01 μM; blue, 0.1 μM; green, 1 μM; orange, 10 μM. The initial value of [P]t is 0.1 mM.

JGGMB2_2018_v22n4_125_f0003.png 이미지

Figure 3. Two-site binding model: two proteins and one ligand. (A) The simulated titration curves for P with L as a function of [L]t/[P]t ratio using Eq. 44 (a: Kd,2 = 1 μM and various Kd,1; b: Kd,1 = 1 μM and various Kd,2). The initial value of [P]tis 0.1 mM. (B) The calculated 1H/15N-HSQC cross-peaks of P during titration with L using (a) Kd,2 of 1 μM and various Kd,1 values and (a) Kd,1 of 1 μM and various Kd,2 values. The cross-peak color changes gradually from blue to purple according to the [L]t/[P]t ratio. The black cross-peak indicates the two bound states (PL and P2L).

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