Modern Solid State NMR. Stefan Steuernagel Bruker BioSpin GmbH, Germany BeNeLux Users Meeting, Brussels November 25, 2016

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1 Modern Solid State NMR Stefan Steuernagel Bruker BioSpin GmbH, Germany BeNeLux Users Meeting, Brussels November 25, 2016

2 Modern Solid State NMR Solid state DNP NMR 111 khz MAS with 0.7mm probe TopSolids 2.0 competence made easy

3 Solid State DNP NMR Many applications in solid state NMR suffer from insufficient sensitivity Solid state DNP NMR today is the method of choice to solve this Biological systems (membrane proteins, fibrils etc.) Pharmaceuticals ( 13 C and 15 N 2D correlations at natural abundance Materials (frameworks, MOFs)

4 DNP of MOFs DNP-enhanced 15 N{ 1 H} CPMAS NMR spectra of UiO-66-NH 2 (a), UiO-66- NH 2 exposed to KCl (b), and Pt 2+ - loaded samples The NH 2 Pt peak grows linearly with Pt loading of the MOF. Estimates of the ratio of free NH 2 vs. NH 2 Pt can be made based on the relative peak intensities in the NMR spectra.

5 Solid-State DNP NMR Spectrometer Gyrotron and Klystron Options DNP with Gyrotron Work in Progress: 263 GHz Klystron Offer a lower-price more compact microwave source at 263 GHz Design compatible with possible upgrade later to gyrotron for full power performance Power output ~ 5 W DNP with Klystron (EIK) Available 2017

6 111kHz - 0.7mm MAS Probe Highest available MAS rate High RF excitation in the 0.5MHz regime MAS stability of better than 0.25 at 111kHz (±28Hz max) 500nl Volume (350nl active volume) Available for SB/WB in HX / HXY / HCN configuration MAS 3 is mandatory!

7 MAS 3 High Accuracy MAS regulation for 0.7 mm Standard deviation less than 4Hz Long term stability for more than 7 hours tested

8 Tools for 0.7 mm rotor handling

9 Solution State / Solid State MAS NMR

10 BL 0.7 mm first results hnh 2D 111kHz Coherence Lifetimes & Linewidths (Gb1) Data courtesy of Guido Pintacuda, ENS Lyon

11 Structure Elucidation of a capsid protein GB1 proved possibilities In 2016 unknown structures are targeted Picture shows a e.g.capsid Data on the following slides courtesy of Guido Pintacuda, ENS Lyon PNAS (33) E4757-E4758

12 1. Backbone Resonance Assignment

13 2. Elucidating >100kHz MAS 15 N-Linked Ha Detected Walk 13 CO Linked Ha Detected Walk -> linking over prolines

14 15 N-Linked Ha Detected Walk

15 3. Rapid Side Chain Assignment

16 4. Proximities Between Protons in Side-Chain

17 5. Automatic Structure Calculation with UNIO (T. Herrmann)

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19 With TopSolids facilitates the delicate setup of solid state NMR experiments by reliable automated procedures whatever your background and experience may be 2 nd generation available with TopSpin free of charge compatible with SB/WB 4 mm and 3.2 mm double and triple channel probes (more to come)

20 TopSolids 2.0 News & Updates different modules in flow bar design: New Preparation (incl. advanced user interface to fetch general information in one step, import of former data possible) Probe Setup (not only for general setup, but for e.g. testing a probe after repair, etc.) Bio (incl. setup and optimization, up to 4D 13 C-detected experiments) New Materials (1D/ 2D X, HX CP and MQMAS experiments) modules use an optimum of automation minimized user interactions

21 Preparation Module New start new user interface if unknown: select the currently installed probe open the new user interface and create your project

22 New User Interface for Project Handling General Information simplified creation of a new project 1 2 TopSolids organizes the data set handling automatically mandatory: fill in directory and sample names select the TopSolids module you want to launch 3

23 New User Interface for Project Handling General Information recommended, but not mandatory: load a shim file 4

24 New User Interface for Project Handling General Information this might take some seconds 4 5 creates the new directory within the active data path creates a parameter data bank for the new project within this directory loads the respective first data set (dependent on the chosen module) opens the next tab automatically, once finished

25 New User Interface for Project Handling Import Previous Data possibility to import a previous TopSolids parameter data bank requirement: New part and serial Nos. of the probe need to match

26 New Parameter Handling TopSolids parameter handling is based on: 1. parameter sets 2. PROSOL entries New 3. parameter data bank ensures reasonable values for parameters at any time whether optimized or not yet at the end of any optimization, the determined parameters are stored in the parameter data bank ready for further usage parameters can be reviewed in form of a PDF report at any time, e.g. to assist fast publication

27 Probe Setup Module from time to time probe performance needs to be tested though routine work, it can be time consuming even for an NMR expert Probe Setup module offers a high level of convenience to: adjust the magic angle optimize 1 H and 13 C hard pulses on standard samples shim the probe and calibrate spectra with least user interaction to overcome the necessity for extensive prior knowledge only two samples needed: KBr, adamantane

28 Probe Setup Module Magic Angle Setting New Magic Angle setting is done on KBr ( 79 Br) at 5 khz MAS TopSolids: checks for fixed frequency H/C/N probes to adjust respective parameter sets automatically New New can detect a Cover Module 2, which can be used for MA adjustment at the spectrometer directly if wanted records a reference spectrum and evaluates the quality of the MA setting

29 TopSolids Material Samples New high demand to implement automation and setup support for applications on material samples TopSolids now provides guided routines to: any X nucleus of choice the mounted probe can handle as for protein applications: optimizations on standard samples and/ or sample of interest for: X hard pulse HX CP 1D/ 2D MQMAS

30 Material Applications Standard Sample Setup guides through basic setup routines of 90 degree pulse calibration and CP setup X 90 degree pulse optimization either as a high or a low power pulse, depending on the X nucleus of choice for MQMAS experiments switch to the nd Experiment module, once the 90 degree pulse is known

31 Material Applications nd Experiments guides step by step through the setup process to run a 2D MQMAS experiment by optimizing: 1. the minimum spectral width needed as indirect dimension to reduce 2D measurement time

32 Material Applications nd Experiments guides step by step through the setup process to run a 2D MQMAS experiment by optimizing: 2. specific MQMAS excitation and reconversion pulse lengths for optimum sensitivity using automated parameter optimization and B 1 excitation field calculations reconversion excitation

33 Material Applications nd Experiments TopSolids: guides through the 2D setup step by step takes care that a multiple of the phase cycle is used as number of scans automatically synchronizes the optimized spectral width with the MAS rate applies basic processing and shearing user interaction only needed for entering number of scans and relaxation delay

34 TopSolids Summary Simplified user interface Suited for protein and material samples, including quadrupolar nuclei New automation features and minimized user interactions, including basic processing up to 3D s Materials modules include HX CP and MQMAS setup Forward & reverse parameter optimization for protein samples Supports 3.2 and 4 mm double and triple channel probes Extended parameter reporting, including information about nd experiments All multidimensional experiments can be queued basic processing is done as soon as an experiment will have been finished

35 Contact core application people solid-state NMR: Gerhard Althoff (DE) Venita Decker (DE) Sebastian Wegner (DE) Barbara Perrone (CH) Jochem Struppe (US) Fabien Aussenac (F) for DNP + all the other application people world wide! important s: solids@bruker.com (DE/CH/FR/US) solidsww@bruker.com (WW)

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