Abstracts and Conference Planner

Oral presentation abstract:

Oral: TOA - Small Molecules 2
Time: 09:50am - 10:30am, Tuesday, 08:30am - 10:30am, Merrill Hall

NMR Supersequences for the Characterisation of Small Molecules

Tim Claridge1; Eriks Kupce2

1University of Oxford, Oxford, United Kingdom; 2Bruker UK, Coventry, United Kingdom

ABSTRACT:

The structure characterisation of small molecules by NMR spectroscopy continues to represent one of the major applications of this analytical technique and routinely employs well established 2D homonuclear and heteronuclear correlation experiments. Now recognised as the leading techniques, attention has turned to developing experimental methods that allow the faster collection of these data sets. Herein, we describe an approach to data collection we term NOAH (NMR by Ordered Acquisition using 1H-detection) that records multiple 2D data sets nested as modules within a single "supersequence". This requires only a single recovery delay for each series of modules, and so allows for significantly reduced data collection times.

 

Oral: TOB - Instrumentation 1
Time: 08:50am - 09:10am, Tuesday, 08:30am - 10:30am, Chapel

High-Power Solid-State Millimeter-Wave Sources to Enable Advanced EPR and DNP-NMR Measurements

Eric Bryerton1; Steven Retzloff1; Melanie Rosay2; Ivan Sergeyev2; Fabien Aussenac3; Jeffrey Hesler1

1Virginia Diodes, Inc., Charlottesville, VA; 2Bruker BioSpin, Billerica, MA; 3Bruker Biospin, Wissembourg,

ABSTRUCT:

DNP uses a high-power millimeter-wave source to irradiate electron spins and transfer high electron polarization to the nuclear spins, thereby enhancing the NMR signal. This fundamentally reduces the time required to acquire the desired data; making a much broader range of measurements feasible. The critical barrier to the greater use of DNP-NMR systems is the lack of sufficiently powerful and frequency agile solid-state sources. In this work, we have developed and demonstrated an electronically tunable multiplier-based solid-state source with 256 mW output power at 262 GHz. This source was used to measure DNP enhancements of 50 using a 3.2 mm sample diameter probe and 120 using a 1.3 mm diameter sample probe, both at 100 K sample temperature.

 

Oral: WOD - Biosolids
Time: 12:00pm - 12:20pm, Wednesday, 11:00am - 1:00pm, Chapel

Elucidating membrane protein - cholesterol binding using distance measurements and DNP-enhanced solid-state NMR

Matthew Elkins1; Ivan Sergeyev2; Mei Hong1

1MIT, Cambridge, MA; 2Bruker BioSpin, Billerica, MA

ABSTRUCT:

Cholesterol plays a major role in membrane protein function, but direct determination of cholesterol-bound structures of proteins in lipid bilayers has not been shown. We have developed several solid-state NMR techniques and biosynthetic isotope labeling for determining cholesterol-bound structures of membrane proteins. Our approaches include 13C-19F distance measurements to constrain the cholesterol tail position from the protein, biosynthetic 13C labeling of cholesterol and 2D 13C-13C correlation NMR with DNP sensitivity enhancement to identify intermolecular contacts, and 2H NMR to determine cholesterol orientation. Applied to the influenza M2 protein, which binds cholesterol to mediate virus budding, our data led to a structural model of the cholesterol-M2 complex, which gives unique insights into how cholesterol promotes the membrane scission function of M2.

 

Oral: ThOC - in(ex)-vivo 2
Time: 12:00pm - 12:20pm, Thursday, 11:00 am - 1:00pm, Merrill Hall

Inverse or Direct Detect Experiments and Probes: Which are "best" for In-vivo Research of 13C Enriched Organisms?

Monica Bastawrous1; Maryam Tabatabaei-Anaraki1; Ronald Soong1; Wolfgang Bermel2; Hermann Heumann3; Andre J Simpson1

1University of Toronto, Toronto, ON; 2Bruker BioSpin, Rheinstetten, Germany; 3Silantes GmbH, München, Germany

ABSTRUCT:

Recent in-vivo metabolic studies have emphasized the importance of 2D data collection and isotopic enrichment of organisms for metabolite identification. However, it has not been determined which multidimensional NMR experiments, inverse proton detected or 13C detected, offers more metabolite information or if they are complementary to each other and should be run simultaneously. Due to the need of uninterrupted experiments during in-vivo analysis, only one probe can be used during a study. Assuming both experiments need to be run, is it best to run them on an inverse or observe probe? To answer these questions and determine an optimal protocol for in-vivo studies HSQC and HETCOR experiments are compared on TCI and BBO cryoprobes using the keystone species D. magna.

 

Oral: FOB - Materials 3
Time: 08:30am - 09:10am, Friday, 08:30am - 10:30am, Chapel

An atomistic view of catalysts using solid-state NMR

Raynald Giovine1; Julien Trébosc1; Frédérique Pourpoint1; Jean-Paul Amoureux1, 2; Olivier Lafon1, 3

1Univ. Lille, CNRS, ENSCL, UCCS, Lille, France; 2Bruker, Wissembourg, France; 3Institut Univ. de France, Paris, France

ABSTRUCT:

We show how advanced NMR techniques and instruments can provide new insights into the atomic-level structure of catalysts. We have notably introduced in recent years efficient and robust techniques to probe connectivities and proximities involving quadrupolar nuclei, such as <sup>17</sup>O, <sup>27</sup>Al, <sup>67</sup>Zn... These techniques can be combined with high magnetic fields and/or Dynamic Nuclear Polarization in order to improve their resolution and sensitivity. These innovative NMR approaches have notably been employed to observe the structure of the Brønsted acid sites in amorphous silica alumina, the synergy between active sites in Zn-modified zeolites, the oxygen sites at the surface of silica nanoparticles and the functionalization of fibrous silica nanoparticles to form solid base and photocatalysts.

 Poster information abstract:

Poster: PA - Solution and Biomolecular
Poster number: 033, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion
  

The 19F-13C TROSY for studies of proteins and nucleic acids

Gerhard Wagner1; Andras Boeszoermenyi1; Sandeep Chhabra1; Abhinav Dubey2; Vladimir Gelev3; Clemens Anklin4; Helena Kovacs5; Ilya Kuprov6; Koh Takeuchi7; Haribabu Arthanari1

1Harvard Medical School, Boston, MA; 2Dana Farber Cancer Institute, Boston, United States; 3Sofia University, Sofia, Bulgaria; 4Bruker Biospin, Billerica, MA; 5Bruker Biospin, Fällanden, Switzerland; 6University of Southampton, Southampton, United Kingdom; 7AIST, Tokyo, Japan

ABSTRACT:  

Use of 19F NMR has long been of interest for studies of proteins but has not become very popular because of the large fluorine line widths. On the other hand, the large 19F CSA suggested to employ a fluorine TROSY effect. However, it was not obvious which nuclear correlation to use, and which nucleus to detect. Extensive calculations with the SPINACH software predicted the field dependence of the aromatic 19F-13C TROSY effect for aromatic amino acids and also for the bases of nucleic acids. Initial experiments on 19F labeled proteins and nucleic acids were consistent with the predictions. Several pulse schemes were explored and will be presented.

 

Poster: PA - Solution and Biomolecular 
Poster number: 034, All Week, 2:00 - 3:45, Fred Farr Pavilion

Heteronuclear direct detection and multiple receivers to acquire simultaneous snapshots of intrinsically disordered proteins in action

Marco Schiavina1; Maria Grazia Murrali1; Letizia Pontoriero1; Valerio Sainati1; Rainer Kümmerle2; Wolfgang Bermel3; Roberta Pierattelli1Isabella C. Felli1

1CERM University of Florence, Florence, Italy; 2Bruker BioSpin AG, Fällanden, Switzerland; 3Bruker BioSpin GmbH, Rheinstetten, Germany

ABSTRACT:  

Intrinsically disordered proteins (IDPs) as well as intrinsically disordered regions (IDRs) of complex protein machineries have recently been recognized as key players in many cellular functions. The CON experiment has become one of the most useful NMR experiments to get a snapshot of an IDP/IDR in conditions approaching physiological ones. Here we propose several variants of the CON experiment in which, during the recovery delay, a second 2D experiment is acquired, either based on 1H detection (CON//HN) or on 15N detection (CON//btNH, CON//(H)CAN). The possibility to collect simultaneous snapshots of an IDP/IDR through different 2D spectra provides a novel tool to follow post-translational modifications, as well as to study samples of limited lifetime such as cell lysates or whole cells.

 

Poster: PA - Solution and Biomolecular
P
oster number: 090, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion 

Ultra-High Field NMR for biological applications: high resolution NMR at 25.8T / 1.1 GHz

Roberta Pierattelli2; Isabella C. Felli2; Rainer Kuemmerle1

1Bruker BioSpin, Fällanden, Switzerland; 2CERM University of Florence, Florence,

ABSTRACT:  

High resolution NMR magnets beyond 23.5 T / 1.0 GHz require all superconducting hybrid HTS/LTS magnets fulfilling tough requirements (10-9) for both field homogeneity and field stability. We report first results of high resolution NMR applications for intrinsically disordered proteins acquired at 25.8 T / 1.1 GHz demonstrating the achievement of magnet, electronics and probe design for this world record field for all superconducting NMR magnets. 

 

Poster: PA - Solution and Biomolecular
Poster number: 119, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Real Time Insights into Biological Events: In-Cell Processes and Protein-Ligand Interactions

Donna M Baldisseri

Bruker BioSpin Corp., Billerica, MA

Poster information abstract:

Poster: PA - Solution and In vivo/Ex vivo 
Poster number: 137, All Week, 2:00 - 3:45, Fred Farr Pavilion

Ex-vivo  Comprehensive Multiphase NMR of Intact Daphnia magna    

Rajshree Ghosh Biswas1; Blythe Fortier-Mcgill1; Ronald Soong1; Daniel Schmidig2; Peter De Castro2; Stephan Graf2; Sebastian Wegner3; Falko Busse3; Jochem Struppe4; Michael Fey4; Myrna J. Simpson1; Andre J. Simpson1

1Environmental NMR Centre, University of Toronto, Toronto, Canada; 2Bruker Biospin AG, Fällanden, Switzerland; 3Bruker BioSpin GmbH, Rheinstetten, Germany; 4Bruker Corporation, Billerica, USA

ABSTRACT:  

Comprehensive Multiphase NMR (CMP-NMR) is a novel technique used to study different phases (solids, solutions and semisolids) in intact samples. This study illustrates multi-phase spectral editing techniques to follow ex vivo samples of Daphnia magna over various life stages and over different generations. The research demonstrates how spectral editing approaches in combination with multiphase NMR help uncover novel information on biological processes. In summary ex vivo NMR proves to be a very powerful approach to study whole organisms in a comprehensive manner and should provide very complimentary information to in vivo based research.  

 

Poster: PA - Solution and * omics / natural products 
Poster number: 162, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Comprehensive Multiphase NMR: A Powerful Tool in Optimizing Biofuel Production

Paris Ning1; Daniel Lane1; Ronald Soong1; Daniel Schmidig2; Thomas Frei2; Peter De Castro2; Ivan Kovacevic2; Stephan Graf2; Sebastian Wegner3; Falko Busse3; Jochem Struppe4; Michael Fey4; Henry J. Stronks5; Martine Monette5; Hermann Heumann6; Myrna J. Simpson1; Andre J Simpson1

1University of Toronto, Toronto, Canada; 2Bruker Biospin AG (Switzerland), Fällanden, Switzerland; 3Bruker BioSpin, Rheinstetten, Germany; 4Bruker BioSpin, Billerica, MA; 5Bruker Ltd., Milton, Canada; 6Silantes GmbH, München, Germany

ABSTRACT:  

Algae is seen as one of the most important sources of biofuel. Comprehensive Multiphase (CMP) NMR combines all electronics and hardware required to give an overview of all phases. Applying CMP-NMR to algae, post extraction, provides a unique insight into what has been extracted (solution phase), what has been swollen (i.e. potential to be extracted with optimization) and what is inaccessible to subcritical water (i.e. that in the true solid-phase). In turn when applied to different algal strains and extraction procedures, CMP-NMR provides a quick overview as to how effective the approach is as-well as identifying the material that remains and could be extracted with further method optimization.

 

Poster: PA - Solution and Instrumentation 
Poster number: 176, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Improving HTS NMR Coil Pulse Shapes and Receive Bandwidth

Ghoncheh Amouzandeh1; Omid Sanati2; Vijaykumar Ramaswamy3; Nicolas Freytag3; Lawrence Hornak2; Arthur Edison2; William W. Brey1

1National High Magnetic Field Labratory / FSU, Tallahassee, FL; 2University of Georgia, Athens, GA; 3Bruker Biospin AG, Fällanden, Switzerland

ABSTRACT:  

Replacing normal metal NMR coils with high-temperature superconducting (HTS) coils can significantly improve the SNR due to the high Q factor of superconducting resonators. However, the high Q factor will reduce the system bandwidth for both reception and excitation which can be problematic for 13C with a wide chemical shift range. Exciting the whole chemical shift dispersion spectrum requires both strong and short RF pulses. Detecting the FID also requires a short dead-time before losing the NMR signal. The RF properties and the ability of the HTS resonators to accurately generate short pulses for excitation are studied in the time domain. The results show adding a shorted stub into the transmit path will shorten the coil rise and fall times. 

 

Poster: PA - Solution and New or unusual science 
Poster number: 181, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Learning from the Jewellery Industry: CNC Micro-milling for Rapid, Cheap and Background Free NMR Micro-coils

Vincent Moxley-Paquette1; Daniel Lane1; Ronald Soong1; Paris Ning1; Monica Bastawrous1; Maysam Pedram2; Md Aminul Haque Talukder2; Ebrahim Ghafar Zadeh2; Dimitri Zverev3; Richard Martin4; Bob Macpherson5; Mike Vargas5; Falko Busse6; Wolfgang Bermel6; Till Kuehn7; Rainer Kuemmerle7; Danijela Al Adwan-Stojilkovic7; Daniel Schmidig7; Michael Fey8; Henry Stronks9; André Simpson1

1University of Toronto, Toronto, ON; 2York University, North York, ON; 3NSCNC Manufacturing LTD, Port Coquitlam, BC; 4IMicrosolder, Meaford, ON; 5Apogee Steel Fabrication Inc., Mississauga, ON; 6Bruker Biospin GmbH, Rheinstetten, Germany; 7Bruker BioSpin AG, Fällanden, Switzerland; 8Bruker Corporation, Billerica, MA; 9Bruker Canada Ltd, Milton, ON

ABSTRACT:  

There is great interest in miniaturizing NMR detection with the ultimate goal of studying metabolism in individual cells, eggs and tiny organisms. Here we explore the potential of micro-milling directly on Teflon electroplated with copper. Multi-turn spiral coils (40 µm wide copper) and a microstrip are fabricated as examples. Furthermore, micro-milling allows sample holders to be easily constructed at the same time as the coil is made. The microstrip was designed, milled, matched/tuned, and tested on the NMR spectrometer within 24 hrs, demonstrating the considerable potential of micro-machining for rapid prototyping of NMR micro sensors. 

 

Poster: PB - Solids and Small Molecules 
Poster number: 188, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Ultrafast Magic Angle Spinning NMR Investigation of Natural Abundance Pharmaceuticals

 Xingyu Lu1; Chengbin Huang1, 2; Karen C. Thompson1; Yu Tsutsumi4; Jean-Paul Amoureux4; Daniel Skomski1; Haichen Nie1; Wei Xu1; Yongchao Su1, 3

1Pharmaceutical Sciences, Merck & Co., Inc., Kenilworth, NJ; 2School of Pharmacy, University of Wisconsin-Madiso, Madison, WI; 3College of Pharmacy, University of Texas at Austin, Austin, TX; 4Bruker BioSpin, Yokohama, Japan

ABSTRACT:  

Solid-state NMR plays an indispensable role in characterizing solid pharmaceutical dosages for the noninvasive, quantitative and high resolution capabilities. The molecular investigation in multicomponent and natural abundance drug products offers both technical challenges and opportunities for this advanced technique. We aim at advancing proton detected spectroscopy under ultrafast magic angle spinning (MAS) of 60-111 kHz in pharmaceutical sciences. Development of novel multidimensional and heteronuclear correlation methods and their applications in the investigation of quantification of pharmaceutical formulation, drug-polymer interaction, and molecular packing of crystalline and amorphous drug molecules will be presented.

  

Poster: PB - Solids and Biomolecular 
Poster number: 233, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

DNP-MAS NMR for the Structural Characterization of Spider Silk Fibers 

Dillan Stengel1; David Onofrei1; Roshan Guia1; Parmida Saadipour1; Shane Pawsey2; Melanie Rosay2; Bennett Addison1; Gregory Holland1

1San Diego State University, San Diego, CA; 2Bruker Biospin Corporation, Billerica, MA 

ABSTRACT:  

Spider dragline silk is an impressive biological polymer with mechanical properties that rival steel and Kevlar. Dragline silk, the strongest silk spiders make, is spun from silk proteins within the major ampullate gland. These silk proteins are comprised of a repetitive core rich in poly(Ala) runs and glycine-rich regions of Gly-Gly-X, or Gly-Pro-Gly-X-X. Unfortunately, the fibers are produced in low yield and take weeks, sometimes months, to isotope label and collect for solid-state NMR studies. In addition, the solid-state NMR spectra exhibit extremely broad 13C and 15N dimensions further hindering S/N. We therefore examined the potential of DNP-MAS NMR for investigating spider silk, improve S/N and enable more advanced three-dimensional (3D) experiments for structural characterization. 

 

Poster: PB - Solids and New or unusual science 
Poster number: 283, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Protein NMR resonance assignment without spectral analysis: 5D SOlid-state Automated Projection SpectroscopY (SO-APSY)

Henry Orton1; Jan Stanek2, 3; Tobias Schubeis2; Dylan Foucaudeau2; Claire Ollier2; Adrian Draney2; Tanguy Le Marchand2; Diane Cala-De Paepe2; Isabella Felli4; Roberta Pierattelli4; Sebastian Hiller5; Wolfgang Bermel6; Guido Pintacuda2

1Australian National University, Canberra, Australia; 2CRMN Lyon, Lyon, France; 3University of Warsaw, Warsaw, Poland; 4CERM - University of Florence, Florence, Italy; 5Biozentrum, Basel, Switzerland; 6Bruker Biospin, Rheinstetten, Germany

ABSTRACT:  

Narrow proton signals, high sensitivity, and efficient coherence transfersprovided by fast magic-angle spinning at high magnetic fields make automated projection spectroscopy feasible in protein solid-state NMR. We present the first ultra-high dimensional implementation of this approach where 5D peak lists are reconstructed from a number of 2D projections for protein samples of different molecular size and aggregation state, featuring limited dispersion of chemical shifts or inhomogeneous broadenings. The resulting datasets are particularly suitable to automated analysis, yielding rapid and unbiased backbone resonance assignments.

 

Poster: PE - Hyperpolarization and Small Molecules 
Poster number: 344, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Cyclic Coherent Hyperpolarization of Water with p-H2

Eric Breynaert1; Ewoud Vaneeckhaute1; Jean-Max Tyburn2; James Kempf3; Francis Taulelle1; Johan Martens1

1COK-KAT, KU Leuven, Leuven, BELGIUM; 2Bruker Biospin, Rheinstetten, Germany; 3Bruker Biospin, Billerica, MA, US

 

Poster: PE - Hyperpolarization and Biomolecular 
Poster number: 358, All Week, 2:00pm- 3:45pm, Fred Farr Pavilion

Biradical-tagged Phospholipid for Matrix-free DNP MAS NMR of Membrane Proteins   

Daryl Good2; Maxim Voinov1; David Bolton2; Meaghan Ward2; Ivan Sergeev3; Marc Caporini3; Peter Scheffer2; Andy Lo2; Melanie Rosay3; Antonin Marek1; Leonid Brown2; Vladimir Ladizhansky2; Alex I. Smirnov1

1North Carolina State University, Raleigh, NC; 2University of Guelph, Guelph, ON; 3Bruker Biospin Ltd., Billerica, MA

ABSTRACT:  

A novel DNP polarizing agent ToSMTSL-PTE representing a phospholipid bearing a biradical – a TOTAPOL derivative – tethered to the polar head group by a flexible linker was synthesized, characterized, and employed to enhance solid-state NMR signal of a lipid-reconstituted integral membrane protein proteorhodopsin .  The results were compared with DNP enhancements obtained with a conventional sample protocol based on dispersing TOTAPOL in a glassy water-glycerol matrix.  Complementary variable temperature EPR and differential scanning calorimetry experiments indicated possible conformational changes and an inhomogeneous distribution of the lipid-based biradicals in lipid bilayers when mixed at high (20 mol%) concentration for maximum DNP enhancement.  Thus, the latter two factors should be also considered for further development of matrix-free DNP of membrane proteins. 

 

Poster: PE - Hyperpolarization and Materials 
Poster number: 370, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Narrow line and Hybrid BDPA-Nitroxide Polarizing Agents for High-Field and Fast-MAS DNP   

Dorothea Wisser1; Ganesan Karthikeyan3; Alicia Lund1; Gilles Casano3; Hakim Karoui3; Maxim Yulikov4; Georges Menzildjian1; Arthur Pinon2; Armin Purea5; Frank Engelke5; Sachin Chaudhari1; Dominik Kubicki2; Aaron Rossini2; Ilia Moroz4; David Gajan1; Christophe Copéret4; Gunnar Jeschke4; Lyndon Emsley2; Anne Lesage1; Olivier Ouari3; Moreno Lelli6

1ENS Lyon, Lyon, France; 2EPFL, Lausanne, Switzerland; 3AixMarseille Université, CNRS, ICR,, Marseille, France; 4ETH Zürich, Zürich, Switzerland; 5Bruker BioSpin GmbH, Rheinstetten, Germany; 6University of Florence, Sesto Fiorentino (Florence), Italy

ABSTRACT:  

DNP can boost sensitivity of MAS NMR by around 2 orders of magnitude at 9.4T. Nevertheless, dinitroxide biradicals suffer for a significant drop in performance at high field and fast MAS. Here we show how these limits can be overcome with new polarizing agents, based on the narrow EPR line BDPA radical. Overhauser DNP of BDPA radical in OTP yields enhancements over 100 at 18.8 T and 40 kHz MAS that increase with MAS and with no depolarization or quenching effects. Hybrid BDPA-nitroxide biradicals on the other hand show the highest enhancements and overall sensitivity gains observed up to now at high field. The potential of these polarizing agents for the characterization of pharmaceutical and functionalized material surfaces is illustrated.

 

Poster: PE - Hyperpolarization and Instrumentation
Poster number: 391, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Microwave field simulations at 263GHz–527GHz for small-diameter MAS-DNP probes focusing the incident microwave beam by means of a waveguide coupler

Armin Purea1; Christian Reiter1; Fabien Aussenac2; Frank Engelke1

1Bruker BioSpin, Rheinstetten, Germany; 2Bruker Biospin, Wissembourg, France

 

 Poster information abstract:

Poster: PA - Solution and Biomolecular
Poster number: 033, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion
  

The 19F-13C TROSY for studies of proteins and nucleic acids

Gerhard Wagner1; Andras Boeszoermenyi1; Sandeep Chhabra1; Abhinav Dubey2; Vladimir Gelev3; Clemens Anklin4; Helena Kovacs5; Ilya Kuprov6; Koh Takeuchi7; Haribabu Arthanari1

1Harvard Medical School, Boston, MA; 2Dana Farber Cancer Institute, Boston, United States; 3Sofia University, Sofia, Bulgaria; 4Bruker Biospin, Billerica, MA; 5Bruker Biospin, Fällanden, Switzerland; 6University of Southampton, Southampton, United Kingdom; 7AIST, Tokyo, Japan

ABSTRACT:  

Use of 19F NMR has long been of interest for studies of proteins but has not become very popular because of the large fluorine line widths. On the other hand, the large 19F CSA suggested to employ a fluorine TROSY effect. However, it was not obvious which nuclear correlation to use, and which nucleus to detect. Extensive calculations with the SPINACH software predicted the field dependence of the aromatic 19F-13C TROSY effect for aromatic amino acids and also for the bases of nucleic acids. Initial experiments on 19F labeled proteins and nucleic acids were consistent with the predictions. Several pulse schemes were explored and will be presented.

 

Poster: PA - Solution and Biomolecular 
Poster number: 034, All Week, 2:00 - 3:45, Fred Farr Pavilion

Heteronuclear direct detection and multiple receivers to acquire simultaneous snapshots of intrinsically disordered proteins in action

Marco Schiavina1; Maria Grazia Murrali1; Letizia Pontoriero1; Valerio Sainati1; Rainer Kümmerle2; Wolfgang Bermel3; Roberta Pierattelli1Isabella C. Felli1

1CERM University of Florence, Florence, Italy; 2Bruker BioSpin AG, Fällanden, Switzerland; 3Bruker BioSpin GmbH, Rheinstetten, Germany

ABSTRACT:  

Intrinsically disordered proteins (IDPs) as well as intrinsically disordered regions (IDRs) of complex protein machineries have recently been recognized as key players in many cellular functions. The CON experiment has become one of the most useful NMR experiments to get a snapshot of an IDP/IDR in conditions approaching physiological ones. Here we propose several variants of the CON experiment in which, during the recovery delay, a second 2D experiment is acquired, either based on 1H detection (CON//HN) or on 15N detection (CON//btNH, CON//(H)CAN). The possibility to collect simultaneous snapshots of an IDP/IDR through different 2D spectra provides a novel tool to follow post-translational modifications, as well as to study samples of limited lifetime such as cell lysates or whole cells.

 

Poster: PA - Solution and Biomolecular
P
oster number: 090, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion 

Ultra-High Field NMR for biological applications: high resolution NMR at 25.8T / 1.1 GHz

Roberta Pierattelli2; Isabella C. Felli2; Rainer Kuemmerle1

1Bruker BioSpin, Fällanden, Switzerland; 2CERM University of Florence, Florence,

ABSTRACT:  

High resolution NMR magnets beyond 23.5 T / 1.0 GHz require all superconducting hybrid HTS/LTS magnets fulfilling tough requirements (10-9) for both field homogeneity and field stability. We report first results of high resolution NMR applications for intrinsically disordered proteins acquired at 25.8 T / 1.1 GHz demonstrating the achievement of magnet, electronics and probe design for this world record field for all superconducting NMR magnets. 

 

Poster: PA - Solution and Biomolecular
Poster number: 119, All Week, 2:00pm - 3:45pm, Fred Farr Pavilion

Real Time Insights into Biological Events: In-Cell Processes and Protein-Ligand Interactions

Donna M Baldisseri

Bruker BioSpin Corp., Billerica, MA