Vasa Suresh K, Singh Himanshu, Rovó Petra, Linser Rasmus
Department Chemistry and Pharmacy , Ludwig-Maximilians-University Munich , Butenandtstr. 5-13 , 81377 Munich , Germany.
Center for Integrated Protein Science (CiPSM), Munich , Germany.
J Phys Chem Lett. 2018 Mar 15;9(6):1307-1311. doi: 10.1021/acs.jpclett.8b00110. Epub 2018 Mar 2.
Solid-state NMR has been employed for characterization of a broad range of biomacromolecules and supramolecular assemblies. However, because of limitations in sensitivity and resolution, the size of the individual monomeric units has rarely exceeded 15 kDa. As such, enzymes, which are often more complex and comprise long peptide chains, have not been easily accessible, even though manifold desirable information could potentially be provided by solid-state NMR studies. Here, we demonstrate that more than 1200 backbone and side-chain chemical shifts can be reliably assessed from minimal sample quantities for a 29 kDa human enzyme of the carbonic anhydrase family, giving access to its backbone dynamics and intermolecular interactions with a small-molecule inhibitor. The possibility of comprehensive assessment of enzymes in this molecular-weight regime without molecular-tumbling-derived limitations enables the study of residue-specific properties important for their mode of action as well as for pharmacological interference in this and many other enzymes.
固态核磁共振已被用于表征多种生物大分子和超分子组装体。然而，由于灵敏度和分辨率的限制，单个单体单元的大小很少超过15 kDa。因此，尽管固态核磁共振研究可能会提供许多有用的信息，但通常更为复杂且包含长肽链的酶却难以进行研究。在此，我们证明，对于碳酸酐酶家族的一种29 kDa人类酶，从极少量样品中就能可靠地评估出1200多个主链和侧链化学位移，从而了解其主链动力学以及与小分子抑制剂的分子间相互作用。在这种分子量范围内对酶进行全面评估而不受分子翻滚限制的可能性，使得研究对其作用模式以及对该酶和许多其他酶的药理干扰很重要的残基特异性性质成为可能。
