Effects of Chemical Exchange on NMR Spectra
|
|
- Winfred Gilbert
- 6 years ago
- Views:
Transcription
1 Effects of Chemical Exchange on NMR Spectra Chemical exchange refers to any process in which a nucleus exchanges between two or more environments in which its NMR parameters (e.g. chemical shift, scalar coupling, or relaxation) differ. DNMR deals with the effects in a broad sense of chemical exchange processes on NMR spectra; and conversely with the information about the changes in the environment of magnetic nuclei that can be derived from observation of NMR spectra. K ex Conformational equilibrium K B Chemical equilibrium
2 Types of Chemical Exchange Intramolecular exchange Motions of sidechains in proteins A B Helix-coil transitions of nucleic acids Unfolding of proteins Conformational equilibria Intermolecular exchange Binding of small molecules to macromolecules Protonation/deprotonation equilibria Isotope exchange processes Enzyme catalyzed reactions M+L ML Because NMR detects the molecular motion itself, rather the numbers of molecules in different states, NMR is able to detect chemical exchange even when the system is in equilibrium
3 2-state First Order Exchange A B k1 k -1 Lifetime of state A: τ A = 1/k +1 Lifetime of state B: τ B = 1/k -1 Use a single lifetime 1/ τ =1/ τ A + 1/τ B = k +1 + k -1
4 Rationale for Chemical Exchange For slow exchange For fast exchange FT FT Bloch equation approach: dm AX /dt = -( ω A )M AY -M AX /τ A + M BX /τ B dm BX /dt = -( ω B )M BY -M BX /τ B + M AX /τ A
5 2-state 2nd Order Exchange k +1 M+L ML k -1 K d = [M] [L]/[ML] = k -1 /k +1 K d = M k on = k +1 ~ 10 8 M -1 s -1 (diffusion-limited) k -1 ~ s -1 Lifetime 1/ τ =1/ τ ML + 1/τ l = k -1 (1+f ML /f L ) f ML and f L are the mole fractions of bound and free ligand, respectively
6 Typical Motion Time Scale for Physical Processes SLOW very slow slow fast very fast ultrafast s ms µs ns ps fs FAST MACROSCOPIC DIFFUSION, FLOW CHEMICAL EXCHANGE MOLECULAR ROTATIONS RELAXATION TIMESCALE SPECTRAL TIMESCALE LARMOR TIMESCALE MOLECULAR VIBRATIONS
7 NMR Time Scale Time Scale Chem. Shift, δ Coupling Const., J T2 relaxation Slow k << δ Α δ Β k << J Α J Β k << 1/ T 2, Α 1/ T 2, B Intermediate k = δ Α δ Β k = J Α J Β k = 1/ T 2, Α 1/ T 2, B Fast k >> δ Α δ Β k >> J Α J Β k >> 1/ T 2, Α 1/ T 2, B Sec NMR time-scale refers to the chemical shift timescale. The range of the rate can be studied s -1 for H can be extended to faster rate using 19 F, 13 C and etc.
8 Slow Exchange k << δ A -δ B A B k1 k -1 Separate lines are observed for each state. The exchange rate can be readily measured from the line widths of the resonances Like the apparent spin-spin relaxation rates, 1/T 2i,obs 1/T 2A,obs = 1/T 2A + 1/τ A = 1/T 2A + 1/k 1 1/T 2B,obs = 1/T 2B + 1/τ Β = 1/T 2B + 1/k -1 line width Lw = 1/πT 2 = 1/πT 2 + k 1 /π Each resonance is broadened by Lw = k/π Increasing temperature k increases, line width increases
9 Slow Exchange for M+L ML k 1 k- 1 Separate resonances potentially are observable for both the free and bound states M F, M B, L F, and L B The addition of a ligand to a solution of a protein can be used to determine the stoichiometry of the complex. Once a stoichiometric mole ratio is achieved, peaks from free ligand appear with increasing intensity as the excess of free ligand increases. Obtain spectra over a range of [L]/[M] ratios from 1 to 10
10 Slow Exchange for M+L ML For free form 1/T 2L,obs = 1/T 2L + 1/τ L = 1/T 2L + k -1 f ML /f L 1/T 2M,obs = 1/T 2M + 1/τ Μ = 1/T 2M + k -1 f ML /f M For complex form 1/T 2ML,obs = 1/T 2ML + 1/τ ML = 1/T 2ML + k -1 Measurements of line width during a titration can be used to derive k -1 (k off ). k 1 k- 1
11 19 F spectra of the enzyme-inhibitor complex at various mole ratio of carbonic anhydrase:inhibitor Free inhibitor 1:4 1:3 1:2 1:1 1:0.5 Bound ligand At -6 ppm the broadened peak for the bound ligand is in slow exchange with the peak from free ligand at 0 ppm. The stoichiometry of the complex is 2:1. No signal from the free ligand is visible until more than 2 moles of inhibitor are present.
12 Coalescence Rate For A B equal concentrations, there will be a rate of interchange where the separate lines for two species are no longer discernible The coalescence rate k c = c π δ δ // 2 2 = δ δ δ is the chemical shift difference between the two signals in the unit of Hz. δ depends on the magnetic field
13 Coalescence Temperature Since the rate depends on the G of the inversion, and the G is affected by T, higher temperature will make things go faster. Tc is the temperature at which fast and slow exchange meet. T>Tc, fast exchange T<Tc, slow exchange T we can calculate the G of the process G G = R ** T C * C *[[ ln ln (( T C / C / δ δ ))]] T C
14 Fast Exchange k >> δ A -δ B A B k1 k -1 A single resonance is observed, whose chemical shift is the weight average of the chemical shifts of the two individual states δ obs = f A δ A +f B δ f B, A + f B = 1 For very fast limit 1/T 2,obs = f A /T 2A + f B /T 2B For moderately fast 1/T 2,obs = f A /T 2A + f B /T 2B + f A f B2 4π ( δ AB ) 2 / k -1 Maximal line broadening is observed when f A = f B = 0.5
15 Line Shape Simulation
16 Fast Exchange k >> δ A -δ B k+1 M+L ML k-1 For M δ M,obs = f M δ M +f ML δ ML For L δ L.obs = f L δ L +f ML δ ML 1/T 2,obs = f ML /T 2,ML + f L /T 2,L + f ML f L2 4π (δ ML -δ L ) 2 / k -1 A maximum in the line broadening of ligand or protein resonances occurs during the titration at a mole ratio of approx. ligand:protein 1:3 The dissociation constant for the complex can be obtained by measuring the chemical shift of the ligand resonance at a series of [L].
17 NMR Titration Both T6(CH3) at 1.1 ppm from DNA (A) and L(CNH 2 ) methyl from ligand (B) are in fast exchange. Expanded regions from 300 MHz 1 HNMR spectra from complexes between L(NH 2 ) and d(ggtaatacc) 2 recorded at 10 ºC (Craik)
18 Residues in the linker of Calmodulin underwent sequential phases of conformational change
19 Binding of Inhibitor to Enzyme H I * H I H * k off * k unf * H S * H * k off [protein][ligand] off K d = d = k on [protein- ligand] ligand] k on H S NMR studies are done at mm, making it difficult to determine K d with any accuracy if the binding is very tight (nm K d ).
20 Measuring Binding Constant k+1 M+L ML k-1 δ M,obs = f M δ M +f ML δ ML, The total change in δ of M δ Mo = δ ML - δ M At [L], δ M = δ M,obs - δ M If [M] is fixed, δ M= δ Mo [L]/([L]+K d ) At 0.5 δ Mo, K d = [L] Similar for A - +H + k+1 k-1 AH H=L, K d = K a δ H= δ Ho [H]/([H]+K a ) Inhibitor G-3-P binds to Triosephapte Isomerase -JMB, :319
21 NMR studies of Ca(II) Binding to cntnc [Ca]t/cTnC cntnc is the N-terminal domain of TnC from cardiac muscle with a single calcium binding site II. Peak at 10.3 ppm is G70 at the calcium binding site. Monica et al., 1997
22 Monitoring Calcium-binding by NMR CaM-CD2-III-5G Ca mm mm mm 0 mm fractional change of chemical shifts ppm K d = 144 ± 27 um [Ca(II)], (mm)
23 The Ionization of His in Oxy & DeOxyhemglobin C2H 7.7 ppm 8.7 ppm C4H 7.0 ppm 7.4 ppm Bohr effect: Hb takes up H + on releasing O2 The pka of His-β-146 changes by more than 1pH unit upon ionization due to the stabilization of the charged form by Asp94 in the deoxy structure.
24 Transferred NOE (TRNOE) Cross-relaxation (NOE) between two protons in the bound ligand is transferred to the free molecular by chemical exchange between bound and free ligand. The negative NOEs from the bound state can be detected by carrying NOE experiments on the free ligand in slow exchange, or one the averaged signals from free and bound ligand in fast exchange. irradiation H1 H2 H1 bound L free L H2 protein
25 Transferred NOE (TRNOE) free H1 k bindr k bindf bound H1 k magf k magr k magf k magr free H2 k bindr k bindf bound H2 H1 H2 H1 bound L free L For small ligands such as short peptides, the enhancement from cross-relaxation is almost 0. On the other hand, large molecules such as receptors have very efficient magnetization transfer. If the chemical exchange rate is great enough, magnetization is transferred from the free protons via the bound state. This gives distance information of the bound state. H2 protein
26 Selectively deuterated dihydrofolate reductase in the presence of 2.7 molar equivalents of trimethoprim control Irriadiation st 7.34 ppm Spectrum a-b H6 is close to H2 (or H6 ) in a folded conformation of bound trimethoprim
27 1 H(pm) Superposition of 50 energy minimized structures of FaNaCh-bound FMRFa N-terminus C-terminal amide
28 Ligand binding: Isotope editing NMR techniques exist to separate a mixture of molecules with different isotopic labels. For example, if two proteins are mixed together and one has 15N labels and the other doesn t, it is possible to only observe the amide protons of one or the other (technically it is easier to see the amides of the protein with 15N, but either is possible). This can be extended to mixtures of pools of ligands with proteins where either the ligand is labeled or the protein is labeled. More complicated mixtures can be analyzed by incorporating different types of labels (e.g. 15N on one, 13C on another)
29 Dynamic NMR to Monitor Rapid Protein Folding The intensity as a function of frequency McConnell equation I(ν) - the intensity at frequency ν C o -constants proportional to [protein] tot P N and P D - the populations of N and D The parameters needed to calculate the lineshape are C o, & ν, T 2N, T 2D, k f and k u. The first four parameters are obtained directly from the spectra using Lorentzian line shape simulation of the J-coupled multiplets. k f and k u can be obtained independently by simulating the position and shape of a given resonance. --J. Sandström, 1982
30
Effects of Chemical Exchange on NMR Spectra
Effects of Chemical Exchange on NMR Spectra Chemical exchange refers to any process in which a nucleus exchanges between two or more environments in which its NMR parameters (e.g. chemical shift, scalar
More informationK ex. Conformational equilibrium. equilibrium K B
Effects of Chemical Exchange on NMR Spectra Chemical exchange refers to any yprocess in which a nucleus exchanges between two or more environments in which its NMR parameters (e.g. chemical shift, scalar
More informationChemical Exchange and Ligand Binding
Chemical Exchange and Ligand Binding NMR time scale Fast exchange for binding constants Slow exchange for tight binding Single vs. multiple binding mode Calcium binding process of calcium binding proteins
More informationGeorgia State University
Chemical Exchange and Calcium Signaling Jenny J. Yang Jenny@gsu.edu Chemistry Department Center for Diagnostics & Therapeutics Georgia State University http://www.gsuyanglab.com/research Ligand Protein
More informationLecture #6 Chemical Exchange
Lecture #6 Chemical Exchange Topics Introduction Effects on longitudinal magnetization Effects on transverse magnetization Examples Handouts and Reading assignments Kowalewski, Chapter 13 Levitt, sections
More informationSlow symmetric exchange
Slow symmetric exchange ϕ A k k B t A B There are three things you should notice compared with the Figure on the previous slide: 1) The lines are broader, 2) the intensities are reduced and 3) the peaks
More informationChemical Exchange. Spin-interactions External interactions Magnetic field Bo, RF field B1
Chemical Exchange Spin-interactions External interactions Magnetic field Bo, RF field B1 Internal Interactions Molecular motions Chemical shifts J-coupling Chemical Exchange 1 Outline Motional time scales
More informationIntroduction solution NMR
2 NMR journey Introduction solution NMR Alexandre Bonvin Bijvoet Center for Biomolecular Research with thanks to Dr. Klaartje Houben EMBO Global Exchange course, IHEP, Beijing April 28 - May 5, 20 3 Topics
More informationNMR in Structural Biology
NMR in Structural Biology Exercise session 2 1. a. List 3 NMR observables that report on structure. b. Also indicate whether the information they give is short/medium or long-range, or perhaps all three?
More informationProtein dynamics from NMR Relaxation data
Protein dynamics from NMR Relaxation data Clubb 3/15/17 (S f2 ) ( e ) Nitrogen-15 relaxation ZZ-exchange R 1 = 1/T 1 Longitudinal relaxation (decay back to z-axis) R 2 = 1/T 2 Spin-spin relaxation (dephasing
More informationI690/B680 Structural Bioinformatics Spring Protein Structure Determination by NMR Spectroscopy
I690/B680 Structural Bioinformatics Spring 2006 Protein Structure Determination by NMR Spectroscopy Suggested Reading (1) Van Holde, Johnson, Ho. Principles of Physical Biochemistry, 2 nd Ed., Prentice
More informationBMB/Bi/Ch 173 Winter 2018
BMB/Bi/Ch 173 Winter 2018 Homework Set 8.1 (100 Points) Assigned 2-27-18, due 3-6-18 by 10:30 a.m. TA: Rachael Kuintzle. Office hours: SFL 220, Friday 3/2 4:00-5:00pm and SFL 229, Monday 3/5 4:00-5:30pm.
More information- Basic understandings: - Mapping interactions:
NMR-lecture April 6th, 2009, FMP Berlin Outline: Christian Freund - Basic understandings: Relaxation Chemical exchange - Mapping interactions: -Chemical shift mapping (fast exchange) Linewidth analysis
More informationNMR in Medicine and Biology
NMR in Medicine and Biology http://en.wikipedia.org/wiki/nmr_spectroscopy MRI- Magnetic Resonance Imaging (water) In-vivo spectroscopy (metabolites) Solid-state t NMR (large structures) t Solution NMR
More information5th CCPN Matt Crump. Thermodynamic quantities derived from protein dynamics
5th CCPN 2005 -Matt Crump Thermodynamic quantities derived from protein dynamics Relaxation in Liquids (briefly!) The fluctuations of each bond vector can be described in terms of an angular correlation
More informationNMR parameters intensity chemical shift coupling constants 1D 1 H spectra of nucleic acids and proteins
Lecture #2 M230 NMR parameters intensity chemical shift coupling constants Juli Feigon 1D 1 H spectra of nucleic acids and proteins NMR Parameters A. Intensity (area) 1D NMR spectrum: integrated intensity
More informationTITAN: Two-dimensional lineshape analysis
TITAN: Two-dimensional lineshape analysis Chris Waudby Christodoulou Group c.waudby@ucl.ac.uk Andres Ramos Lisa Cabrita John Christodoulou Inhibition of fatty acid synthesis for treatment of tularemia
More informationLineShapeKin NMR Line Shape Analysis Software for Studies of Protein-Ligand Interaction Kinetics
LineShapeKin NMR Line Shape Analysis Software for Studies of Protein-Ligand Interaction Kinetics http://lineshapekin.net Spectral intensity Evgenii L. Kovrigin Department of Biochemistry, Medical College
More informationIntroduction to Relaxation Theory James Keeler
EUROMAR Zürich, 24 Introduction to Relaxation Theory James Keeler University of Cambridge Department of Chemistry What is relaxation? Why might it be interesting? relaxation is the process which drives
More informationSpin-spin coupling I Ravinder Reddy
Spin-spin coupling I Ravinder Reddy Spin-interactions External interactions Magnetic field Bo, RF field B1 Internal Interactions Molecular motions Exchange Chemical shifts J-coupling Spin Diffusion Dipolar
More informationNMR-spectroscopy of proteins in solution. Peter Schmieder
NMR-spectroscopy of proteins in solution Basic aspects of NMR-Spektroskopie Basic aspects of NMR-spectroscopy 3/84 Prerequisite for NMR-spectroscopy is a nuclear spin that can be thought of as a mixture
More informationProteins are not rigid structures: Protein dynamics, conformational variability, and thermodynamic stability
Proteins are not rigid structures: Protein dynamics, conformational variability, and thermodynamic stability Dr. Andrew Lee UNC School of Pharmacy (Div. Chemical Biology and Medicinal Chemistry) UNC Med
More informationBiochemistry 530 NMR Theory and Practice
Biochemistry 530 NMR Theory and Practice Gabriele Varani Department of Biochemistry and Department of Chemistry University of Washington Lecturer: Gabriele Varani Biochemistry and Chemistry Room J479 and
More informationBiophysical Chemistry: NMR Spectroscopy
Relaxation & Multidimensional Spectrocopy Vrije Universiteit Brussel 9th December 2011 Outline 1 Relaxation 2 Principles 3 Outline 1 Relaxation 2 Principles 3 Establishment of Thermal Equilibrium As previously
More informationBiochemistry 530 NMR Theory and Practice. Gabriele Varani Department of Biochemistry and Department of Chemistry University of Washington
Biochemistry 530 NMR Theory and Practice Gabriele Varani Department of Biochemistry and Department of Chemistry University of Washington 1D spectra contain structural information.. but is hard to extract:
More informationPrinciples of Physical Biochemistry
Principles of Physical Biochemistry Kensal E. van Hold e W. Curtis Johnso n P. Shing Ho Preface x i PART 1 MACROMOLECULAR STRUCTURE AND DYNAMICS 1 1 Biological Macromolecules 2 1.1 General Principles
More informationT 1, T 2, NOE (reminder)
T 1, T 2, NOE (reminder) T 1 is the time constant for longitudinal relaxation - the process of re-establishing the Boltzmann distribution of the energy level populations of the system following perturbation
More informationChapter 7. Nuclear Magnetic Resonance Spectroscopy
Chapter 7 Nuclear Magnetic Resonance Spectroscopy I. Introduction 1924, W. Pauli proposed that certain atomic nuclei have spin and magnetic moment and exposure to magnetic field would lead to energy level
More informationQuantification of Dynamics in the Solid-State
Bernd Reif Quantification of Dynamics in the Solid-State Technische Universität München Helmholtz-Zentrum München Biomolecular Solid-State NMR Winter School Stowe, VT January 0-5, 206 Motivation. Solid
More informationNMR-spectroscopy in solution - an introduction. Peter Schmieder
NMR-spectroscopy in solution - an introduction 2/92 Advanced Bioanalytics NMR-Spectroscopy Introductory session (11:00 12:30) Basic aspects of NMR-spectroscopy NMR parameter Multidimensional NMR-spectroscopy
More informationTimescales of Protein Dynamics
Timescales of Protein Dynamics From Henzler-Wildman and Kern, Nature 2007 Dynamics from NMR Show spies Amide Nitrogen Spies Report On Conformational Dynamics Amide Hydrogen Transverse Relaxation Ensemble
More informationProtein-protein interactions (PPIs) via NMR. Paola Turano
Protein-protein interactions (PPIs) via NMR Paola Turano turano@cerm.unifi.it The magnetic field at the The chemical shift nucleus (the effective field) is generally less than the applied field by a fraction
More informationTimescales of Protein Dynamics
Timescales of Protein Dynamics From Henzler-Wildman and Kern, Nature 2007 Summary of 1D Experiment time domain data Fourier Transform (FT) frequency domain data or Transverse Relaxation Ensemble of Nuclear
More informationProtein-protein interactions (PPIs) via NMR. Paola Turano
Protein-protein interactions (PPIs) via NMR Paola Turano turano@cerm.unifi.it The magnetic field at the The chemical shift nucleus (the effective field) is generally less than the applied field by a fraction
More informationSupplementary Information. Overlap between folding and functional energy landscapes for. adenylate kinase conformational change
Supplementary Information Overlap between folding and functional energy landscapes for adenylate kinase conformational change by Ulrika Olsson & Magnus Wolf-Watz Contents: 1. Supplementary Note 2. Supplementary
More informationName: BCMB/CHEM 8190, BIOMOLECULAR NMR FINAL EXAM-5/5/10
Name: BCMB/CHEM 8190, BIOMOLECULAR NMR FINAL EXAM-5/5/10 Instructions: This is an open book, limited time, exam. You may use notes you have from class and any text book you find useful. You may also use
More informationFiltered/edited NOESY spectra
Filtered/edited NOESY spectra NMR Seminar HS 207 Nina Ripin 22..7 Overview NMR of biomolecular complexes Problems and Solutions Filtered/edited nomenclature Experimental elements NOESY vs filtered pulse
More informationBiochemistry 530 NMR Theory and Practice
Biochemistry 530 NMR Theory and Practice Gabriele Varani Department of Biochemistry and Department of Chemistry University of Washington 1D spectra contain structural information.. but is hard to extract:
More informationNMR Spectroscopy. Guangjin Hou
NMR Spectroscopy Guangjin Hou 22-04-2009 NMR History 1 H NMR spectra of water H NMR spectra of water (First NMR Spectra on Water, 1946) 1 H NMR spectra ethanol (First bservation of the Chemical Shift,
More informationNMR Spectroscopy: A Quantum Phenomena
NMR Spectroscopy: A Quantum Phenomena Pascale Legault Département de Biochimie Université de Montréal Outline 1) Energy Diagrams and Vector Diagrams 2) Simple 1D Spectra 3) Beyond Simple 1D Spectra 4)
More informationPRACTICAL ASPECTS OF NMR RELAXATION STUDIES OF BIOMOLECULAR DYNAMICS
PRACTICAL ASPECTS OF MR RELAXATIO STUDIES OF BIOMOLECULAR DYAMICS Further reading: Can be downloaded from my web page Korzhnev D.E., Billeter M., Arseniev A.S., and Orekhov V. Y., MR Studies of Brownian
More informationNMR BMB 173 Lecture 16, February
NMR The Structural Biology Continuum Today s lecture: NMR Lots of slides adapted from Levitt, Spin Dynamics; Creighton, Proteins; And Andy Rawlinson There are three types of particles in the universe Quarks
More informationτ 1 > 1/J - if this lifetime is significantly shortened, the coupling (splitting of the signal) will not be observed
It is often advantageous to reverse or remove the splitting caused by spin-spin coupling This is called spin decoupling Spin decoupling (or just decoupling) can be used for several reasons - to simplify
More informationProtein Dynamics, Allostery and Function
Protein Dynamics, Allostery and Function Lecture 2. Protein Dynamics Xiaolin Cheng UT/ORNL Center for Molecular Biophysics SJTU Summer School 2017 1 Functional Protein Dynamics Proteins are dynamic and
More informationPROTEIN NMR SPECTROSCOPY
List of Figures List of Tables xvii xxvi 1. NMR SPECTROSCOPY 1 1.1 Introduction to NMR Spectroscopy 2 1.2 One Dimensional NMR Spectroscopy 3 1.2.1 Classical Description of NMR Spectroscopy 3 1.2.2 Nuclear
More informationNMR journey. Introduction to solution NMR. Alexandre Bonvin. Topics. Why use NMR...? Bijvoet Center for Biomolecular Research
2 NMR journey Introduction to solution NMR Alexandre Bonvin Bijvoet Center for Biomolecular Research with thanks to Dr. Klaartje Houben EMBO Global Exchange course, CCMB, Hyderabad, India November 29th
More informationInteractions and Dynamics within the Troponin Complex
Interactions and Dynamics within the Troponin Complex Tharin Blumenschein Steve Matthews Lab - Imperial College London (formerly Brian Sykes Lab, Canada) Striated muscle Thin filament proteins - regulation
More informationVIII Chemical Exchange
VIII Chemical Exchange Lecture notes by Assaf Tal Chemical exchange has surprising ties with relaxation as we shall see. Understanding exchange lets us understand phenomena, some of which at first glance
More informationMagnetic Nuclei other than 1 H
Magnetic Nuclei other than 1 H 2 H (Deuterium): I = 1 H,D-Exchange might be used to simplify 1 H-NMR spectra since H-D couplings are generally small; - - - -O- - - -D 2 -O- triplet of triplets slightly
More informationBiophysical Chemistry: NMR Spectroscopy
Spin Dynamics & Vrije Universiteit Brussel 25th November 2011 Outline 1 Pulse/Fourier Transform NMR Thermal Equilibrium Effect of RF Pulses The Fourier Transform 2 Symmetric Exchange Between Two Sites
More informationCHEM / BCMB 4190/6190/8189. Introductory NMR. Lecture 10
CHEM / BCMB 490/690/889 Introductory NMR Lecture 0 - - CHEM 490/690 Spin-Echo The spin-echo pulse sequence: 90 - τ - 80 - τ(echo) Spins echoes are widely used as part of larger pulse sequence to refocus
More informationSequential resonance assignments in (small) proteins: homonuclear method 2º structure determination
Lecture 9 M230 Feigon Sequential resonance assignments in (small) proteins: homonuclear method 2º structure determination Reading resources v Roberts NMR of Macromolecules, Chap 4 by Christina Redfield
More informationNMR Spectroscopy of Polymers
UNESCO/IUPAC Course 2005/2006 Jiri Brus NMR Spectroscopy of Polymers Brus J 1. part At the very beginning the phenomenon of nuclear spin resonance was studied predominantly by physicists and the application
More informationSpin Relaxation and NOEs BCMB/CHEM 8190
Spin Relaxation and NOEs BCMB/CHEM 8190 T 1, T 2 (reminder), NOE T 1 is the time constant for longitudinal relaxation - the process of re-establishing the Boltzmann distribution of the energy level populations
More informationMeasuring Spin-Lattice Relaxation Time
WJP, PHY381 (2009) Wabash Journal of Physics v4.0, p.1 Measuring Spin-Lattice Relaxation Time L.W. Lupinski, R. Paudel, and M.J. Madsen Department of Physics, Wabash College, Crawfordsville, IN 47933 (Dated:
More informationTopics in SSNMR and Dynamics of Proteins: Consequences of Intermediate Exchange
Topics in SSNMR and Dynamics of Proteins: Consequences of Intermediate Exchange A McDermott, Columbia University Winter School in Biomolecular NMR, Stowe VT January 20-23 2008 Effects on NMR Spectra: Local,
More informationProtein Structure Determination using NMR Spectroscopy. Cesar Trinidad
Protein Structure Determination using NMR Spectroscopy Cesar Trinidad Introduction Protein NMR Involves the analysis and calculation of data collected from multiple NMR techniques Utilizes Nuclear Magnetic
More informationLecture #7 In Vivo Water
Lecture #7 In Vivo Water Topics Hydration layers Tissue relaxation times Magic angle effects Magnetization Transfer Contrast (MTC) CEST Handouts and Reading assignments Mathur-De Vre, R., The NMR studies
More informationSpectral Broadening Mechanisms
Spectral Broadening Mechanisms Lorentzian broadening (Homogeneous) Gaussian broadening (Inhomogeneous, Inertial) Doppler broadening (special case for gas phase) The Fourier Transform NC State University
More informationMOLECULAR SPECTROSCOPY AND PHOTOCHEMISTRY
20 CHAPTER MOLECULAR SPECTROSCOPY AND PHOTOCHEMISTRY 20.1 Introduction to Molecular Spectroscopy 20.2 Experimental Methods in Molecular Spectroscopy 20.3 Rotational and Vibrational Spectroscopy 20.4 Nuclear
More informationAnalysis of NMR Spectra Part 2
Analysis of NMR Spectra Part 2-1- Analysis of NMR Spectra Part 2 "Things should be made as simple as possible, but not any simpler." Albert Einstein 1.1 Review of Basic NMR Concepts NMR analysis is a complex
More informationSpin Dynamics Basics of Nuclear Magnetic Resonance. Malcolm H. Levitt
Spin Dynamics Basics of Nuclear Magnetic Resonance Second edition Malcolm H. Levitt The University of Southampton, UK John Wiley &. Sons, Ltd Preface xxi Preface to the First Edition xxiii Introduction
More informationRelaxation. Ravinder Reddy
Relaxation Ravinder Reddy Relaxation What is nuclear spin relaxation? What causes it? Effect on spectral line width Field dependence Mechanisms Thermal equilibrium ~10-6 spins leads to NMR signal! T1 Spin-lattice
More informationNMR course at the FMP: NMR of organic compounds and small biomolecules - II -
NMR course at the FMP: NMR of organic compounds and small biomolecules - II - 16.03.2009 The program 2/76 CW vs. FT NMR What is a pulse? Vectormodel Water-flip-back 3/76 CW vs. FT CW vs. FT 4/76 Two methods
More informationPolarised Nucleon Targets for Europe, 2nd meeting, Bochum 2005
Polarised Nucleon Targets for Europe, nd meeting, Bochum Temperature dependence of nuclear spin-lattice relaxations in liquid ethanol with dissolved TEMPO radicals H. Štěpánková, J. Englich, J. Kohout,
More informationSubstrate-dependent switching of the allosteric binding mechanism of a dimeric enzyme
Supplementary Information: Substrate-dependent switching of the allosteric binding mechanism of a dimeric enzyme Lee Freiburger, 1 Teresa Miletti, 1 Siqi Zhu, 1 Oliver Baettig, Albert Berghuis, Karine
More informationIntroduction to solution NMR. Alexandre Bonvin. The NMR research group. Bijvoet Center for Biomolecular Research
Introduction to solution NMR 1 Alexandre Bonvin Bijvoet Center for Biomolecular Research with thanks to Dr. Klaartje Houben Bente%Vestergaard% The NMR research group Prof. Marc Baldus Prof. Rolf Boelens
More informationUsing NMR to study Macromolecular Interactions. John Gross, BP204A UCSF. Nov 27, 2017
Using NMR to study Macromolecular Interactions John Gross, BP204A UCSF Nov 27, 2017 Outline Review of basic NMR experiment Multidimensional NMR Monitoring ligand binding Structure Determination Review:
More informationInterpreting and evaluating biological NMR in the literature. Worksheet 1
Interpreting and evaluating biological NMR in the literature Worksheet 1 1D NMR spectra Application of RF pulses of specified lengths and frequencies can make certain nuclei detectable We can selectively
More informationPRACTICAL ASPECTS OF NMR RELAXATION STUDIES OF BIOMOLECULAR DYNAMICS
PRACTICAL ASPECTS OF MR RELAXATIO STUDIES OF BIOMOLECULAR DYAMICS Further reading: (Can be downloaded from my web page Korzhnev D.E., Billeter M., Arseniev A.S., and Orekhov V. Y., MR Studies of Brownian
More informationBasic principles of multidimensional NMR in solution
Basic principles of multidimensional NMR in solution 19.03.2008 The program 2/93 General aspects Basic principles Parameters in NMR spectroscopy Multidimensional NMR-spectroscopy Protein structures NMR-spectra
More informationLecture #6 (The NOE)
Lecture #6 (The OE) 2/18/15 Clubb Determining Protein tructures by MR: Measure thousands of shorter inter-hydrogen atom distances. Use these to restrain the structure of protein computationally. Distance
More informationMagnetisation Transfer Schemes
Magnetisation Transfer Schemes P. K. Madhu Department of Chemical Sciences Tata Institute of Fundamental Research Homi Bhabha Road Colaba Mumbai 400 005, India Sensitivity of NMR Spectroscopy S/N Nγ exc
More informationThe wonderful world of NUCLEIC ACID NMR!
Lecture 12 M230 Feigon Sequential resonance assignments in DNA (and RNA): homonuclear method 2 structure determination Reading resources Evans Chap 9 The wonderful world of NUCLEIC ACID NMR! Catalytically
More informationNMR Relaxation and Molecular Dynamics
Ecole RMN Cargese Mars 2008 NMR Relaxation and Molecular Dynamics Martin Blackledge IBS Grenoble Carine van Heijenoort ICSN, CNRS Gif-sur-Yvette Solution NMR Timescales for Biomolecular Motion ps ns µs
More informationContents. xiii. Preface v
Contents Preface Chapter 1 Biological Macromolecules 1.1 General PrincipIes 1.1.1 Macrornolecules 1.2 1.1.2 Configuration and Conformation Molecular lnteractions in Macromolecular Structures 1.2.1 Weak
More informationBiochemistry 530 NMR Theory and Practice
Biochemistry 530 NMR Theory and Practice David Baker Autumn Quarter 2014 Slides Courtesy of Gabriele Varani Recommended NMR Textbooks Derome, A. E. (1987) Modern NMR Techniques for Chemistry Research,
More informationBMB/Bi/Ch 173 Winter 2018
BMB/Bi/Ch 173 Winter 2018 Homework Set 8.1 (100 Points) Assigned 2-27-18, due 3-6-18 by 10:30 a.m. TA: Rachael Kuintzle. Office hours: SFL 220, Friday 3/2 4-5pm and SFL 229, Monday 3/5 4-5:30pm. 1. NMR
More informationNMR Resonance Assignment Assisted by Mass Spectrometry
NMR Resonance Assignment Assisted by Mass Spectrometry This lecture talked about a NMR resonance assignment assisted by mass spectrometry [1, 2]. 1 Motivation Nuclear magnetic resonance (NMR) provides
More informationLongitudinal-relaxation enhanced fast-pulsing techniques: New tools for biomolecular NMR spectroscopy
Longitudinal-relaxation enhanced fast-pulsing techniques: New tools for biomolecular NMR spectroscopy Bernhard Brutscher Laboratoire de Résonance Magnétique Nucléaire Institut de Biologie Structurale -
More informationChemistry 431. Lecture 23
Chemistry 431 Lecture 23 Introduction The Larmor Frequency The Bloch Equations Measuring T 1 : Inversion Recovery Measuring T 2 : the Spin Echo NC State University NMR spectroscopy The Nuclear Magnetic
More informationChem 325 NMR Intro. The Electromagnetic Spectrum. Physical properties, chemical properties, formulas Shedding real light on molecular structure:
Physical properties, chemical properties, formulas Shedding real light on molecular structure: Wavelength Frequency ν Wavelength λ Frequency ν Velocity c = 2.998 10 8 m s -1 The Electromagnetic Spectrum
More informationProtein-protein interactions by NMR
Protein-protein interactions by NMR Fast k on,off >> (ν free - ν bound ) A + B k on k off AB k on,off ~ (ν free - ν bound ) Slow k on,off
More information3 Chemical exchange and the McConnell Equations
3 Chemical exchange and the McConnell Equations NMR is a technique which is well suited to study dynamic processes, such as the rates of chemical reactions. The time window which can be investigated in
More information1. 3-hour Open book exam. No discussion among yourselves.
Lecture 13 Review 1. 3-hour Open book exam. No discussion among yourselves. 2. Simple calculations. 3. Terminologies. 4. Decriptive questions. 5. Analyze a pulse program using density matrix approach (omonuclear
More informationNMR study of complexes between low molecular mass inhibitors and the West Nile virus NS2B-NS3 protease
University of Wollongong Research Online Faculty of Science - Papers (Archive) Faculty of Science, Medicine and Health 2009 NMR study of complexes between low molecular mass inhibitors and the West Nile
More informationChristopher Pavlik Bioanalytical Chemistry March 2, 2011
Nuclear Magnetic Resonance of Proteins Christopher Pavlik Bioanalytical Chemistry March 2, 2011 Nuclear Magnetic Resonance NMR Application of a magnetic field causes absorption of EM energy that induces
More information10.4 Continuous Wave NMR Instrumentation
10.4 Continuous Wave NMR Instrumentation coherent detection bulk magnetization the rotating frame, and effective magnetic field generating a rotating frame, and precession in the laboratory frame spin-lattice
More informationSensitive NMR Approach for Determining the Binding Mode of Tightly Binding Ligand Molecules to Protein Targets
Supporting information Sensitive NMR Approach for Determining the Binding Mode of Tightly Binding Ligand Molecules to Protein Targets Wan-Na Chen, Christoph Nitsche, Kala Bharath Pilla, Bim Graham, Thomas
More informationA Hands on Introduction to NMR Lecture #1 Nuclear Spin and Magnetic Resonance
A Hands on Introduction to NMR 22.920 Lecture #1 Nuclear Spin and Magnetic Resonance Introduction - The aim of this short course is to present a physical picture of the basic principles of Nuclear Magnetic
More informationThe Positive Muon as a Probe in Chemistry. Dr. Iain McKenzie ISIS Neutron and Muon Source STFC Rutherford Appleton Laboratory
The Positive Muon as a Probe in Chemistry Dr. Iain McKenzie ISIS Neutron and Muon Source STFC Rutherford Appleton Laboratory I.McKenzie@rl.ac.uk µsr and Chemistry Properties of atoms or molecules containing
More information1) NMR is a method of chemical analysis. (Who uses NMR in this way?) 2) NMR is used as a method for medical imaging. (called MRI )
Uses of NMR: 1) NMR is a method of chemical analysis. (Who uses NMR in this way?) 2) NMR is used as a method for medical imaging. (called MRI ) 3) NMR is used as a method for determining of protein, DNA,
More information8.2 The Nuclear Overhauser Effect
8.2 The Nuclear Overhauser Effect Copyright Hans J. Reich 2016 All Rights Reserved University of Wisconsin An important consequence of DD relaxation is the Nuclear Overhauser Effect, which can be used
More informationNMR SATELLITES AS A PROBE FOR CHEMICAL
NMR SATELLITES AS A PROBE FOR CHEMICAL INVESTIGATIONS SHIzuo FUJIWARA, Yogi ARATA, HIR0sHI OZAWA and MASAYUKI KuruGI Department of Chemistry, The University of Tokyo, Japan ABSTRACT Satellite lines in
More informationProton NMR. Four Questions
Proton NMR Four Questions How many signals? Equivalence Where on spectrum? Chemical Shift How big? Integration Shape? Splitting (coupling) 1 Proton NMR Shifts Basic Correlation Chart How many 1 H signals?
More informationProtein Dynamics, Allostery and Function
Protein Dynamics, Allostery and Function Lecture 3. Protein Dynamics Xiaolin Cheng UT/ORNL Center for Molecular Biophysics SJTU Summer School 2017 1 Obtaining Dynamic Information Experimental Approaches
More informationBiochemistry 3100 Sample Problems Binding proteins, Kinetics & Catalysis
(1) Draw an approximate denaturation curve for a typical blood protein (eg myoglobin) as a function of ph. (2) Myoglobin is a simple, single subunit binding protein that has an oxygen storage function
More informatione 2m p c I, (22.1) = g N β p I(I +1), (22.2) = erg/gauss. (22.3)
Chemistry 26 Molecular Spectra & Molecular Structure Week # 7 Nuclear Magnetic Resonance Spectroscopy Along with infrared spectroscopy, nuclear magnetic resonance (NMR) is the most important method available
More informationPhysikalische Chemie IV (Magnetische Resonanz) HS Solution Set 2. Hand out: Hand in:
Solution Set Hand out:.. Hand in:.. Repetition. The magnetization moves adiabatically during the application of an r.f. pulse if it is always aligned along the effective field axis. This behaviour is observed
More informationUNIVERSITY OF CINCINNATI
UNIVERSITY OF CINCINNATI Date: I,, hereby submit this work as part of the requirements for the degree of: in: It is entitled: This work and its defense approved by: Chair: Nuclear Magnetic Resonance Studies
More information