title: On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR.
abstract: A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation.
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15260, USA.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Mandal
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15260, USA.
authors.forename: Jennifer C
authors.identifier: 0000-0002-8240-5778
authors.initials: JC
authors.lastname: Boatz
authors.affiliation: Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, 15260, USA.
authors.forename: Travis B
authors.initials: TB
authors.lastname: Wheeler
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15260, USA. vanderwel@pitt.edu.
authors.forename: Patrick C A
authors.identifier: 0000-0002-5390-3321
authors.initials: PC
authors.lastname: van der Wel
mesh_terms: D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010455:Peptides / Q000737:chemistry; D066329:Protein Aggregates; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure; D014461:Ultracentrifugation / Q000295:instrumentation / Q000379:methods
chemical_list: D008565:Membrane Proteins; D010455:Peptides; D066329:Protein Aggregates; D011506:Proteins
keywords: Hydration; MAS NMR; Sample packing; Sedimentation; Ultracentrifugation