| Summary: | Solution-state distance restraints for protein structure determination with Ångström-level resolution rely on through-space transfer of magnetization between nuclear spins. Such magnetization transfers, named Overhauser effects, occur via dipolar magnetic couplings. We demonstrate improvements in magnetization transfer using long-lived coherences (LLCs)—singlet-triplet superpositions that are antisymmetric with respect to spin-permutation within pairs of coupled magnetic nuclei—as the magnetization source. Magnetization transfers in the presence of radio-frequency irradiation, known as ‘rotating-frame’ Overhauser effects (ROEs), are predicted by theory to improve by the use of LLCs; calculations are matched by preliminary experiments herein. The LLC-ROE transfers were compared to the transmission of magnetization via classical transverse routes. Long-lived coherences accumulate magnetization on an external third proton, K, with transfer rates that depended on the tumbling regime. <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mfenced close="}" open="{"><mrow><mi>I</mi><mo>,</mo><mi>S</mi></mrow></mfenced><mo> </mo><mo>→</mo><mi>K</mi></mrow></semantics></math></inline-formula> transfers in the LLC configuration for (I,S) are anticipated to match, and then overcome, the same transfer rates in the classical configuration as the molecular rotational correlation times increase. Experimentally, we measured the LLC-ROE transfer in dipeptide AlaGly between aliphatic protons in different residues <i>K</i> = <i>Ala</i> − <i>H<sup>α</sup></i> and (<i>I</i>,<i>S</i>) <i>= Gly</i> − <i>H<sup>α</sup></i><sup>1,2</sup> over a distance <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mi>d</mi><mfenced close="]" open="["><mrow><mi>K</mi><mo>,</mo><mfenced><mrow><mi>I</mi><mo>,</mo><mi>S</mi></mrow></mfenced></mrow></mfenced></mrow></semantics></math></inline-formula> = 2.3 Å. Based on spin dynamics calculations, we anticipate that, for such distances, a superior transfer of magnetization occurs using LLC-ROE compared to classical ROE at correlation times above <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><msub><mi>τ</mi><mi>C</mi></msub><mo>=</mo><mn>10</mn><mo> </mo><mi>n</mi><mi>s</mi></mrow></semantics></math></inline-formula>. The LLC-ROE effect shows potential for improving structural studies of large proteins and offering constraints of increased precision for high-affinity protein-ligand complexes in slow tumbling in the liquid state.
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