Scholarlink

<p>The nuclear spin-lattice relaxation rate 1/<i>T</i><sub>1</sub> depends on the correlation time <i>τ</i><sub><i>c</i></sub> of the molecule bearing the nuclear spin, and can therefore probe changes of <i>τ</i><sub><i>c</i></sub> upon binding of a rapidly moving small ligand to a more slowly moving larger protein. In practice however, the dependence is such that only a small difference in relaxation rate is obtained at high field. Here we present a scheme in which nuclear spins are first hyperpolarized using DNP, and then allowed to relax at low magnetic field in presence of a target protein, which generates a large <i>T</i><sub>1</sub> contrast. The sample is subsequently transferred into a conventional nuclear magnetic resonance probe (NMR), where the effect of the low-field relaxation is read out using high-field liquid-state NMR. Using only 14 μM of a <sup>13</sup>C-labeled reporter ligand, we observe protein binding reliably for protein concentrations as low as 2 μM in a single scan. The scheme is expanded to a label-free ligand via a competitive binding experiment in which the label-free ligand displaces the <sup>13</sup>C-labeled reporter ligand.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Single-scan detection of ligand-binding using hyperpolarization and low-field relaxation

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年2月21日

查看全文
  • Pooja Narwal,
  • Nils Lorz,
  • Masoud Minaei,
  • Alvar D. Gossert,
  • Benno Meier

摘要

The nuclear spin-lattice relaxation rate 1/T1 depends on the correlation time τc of the molecule bearing the nuclear spin, and can therefore probe changes of τc upon binding of a rapidly moving small ligand to a more slowly moving larger protein. In practice however, the dependence is such that only a small difference in relaxation rate is obtained at high field. Here we present a scheme in which nuclear spins are first hyperpolarized using DNP, and then allowed to relax at low magnetic field in presence of a target protein, which generates a large T1 contrast. The sample is subsequently transferred into a conventional nuclear magnetic resonance probe (NMR), where the effect of the low-field relaxation is read out using high-field liquid-state NMR. Using only 14 μM of a 13C-labeled reporter ligand, we observe protein binding reliably for protein concentrations as low as 2 μM in a single scan. The scheme is expanded to a label-free ligand via a competitive binding experiment in which the label-free ligand displaces the 13C-labeled reporter ligand.