Volume 21, Issue 1, Pages 9-19 (January 2013)

Slides:



Advertisements
Similar presentations
Probing α-310 Transitions in a Voltage-Sensing S4 Helix
Advertisements

Po-chia Chen, Jochen S. Hub  Biophysical Journal 
Networks of Dynamic Allostery Regulate Enzyme Function
Probing α-310 Transitions in a Voltage-Sensing S4 Helix
Volume 18, Issue 1, Pages (January 2011)
Conformational Heterogeneity in the Activation Mechanism of Bax
Volume 23, Issue 9, Pages (September 2015)
Volume 17, Issue 2, Pages (February 2009)
Chaoyou Xue, Natalie R. Whitis, Dipali G. Sashital  Molecular Cell 
Fluorescence Applications in Molecular Neurobiology
Volume 21, Issue 1, Pages (January 2013)
Sudha Chakrapani, Luis G. Cuello, D. Marien Cortes, Eduardo Perozo 
Volume 14, Issue 9, Pages (September 2006)
The Binding of Antibiotics in OmpF Porin
Volume 17, Issue 12, Pages (December 2009)
Complementary Structural Mass Spectrometry Techniques Reveal Local Dynamics in Functionally Important Regions of a Metastable Serpin  Xiaojing Zheng,
Volume 24, Issue 12, Pages (December 2016)
Volume 103, Issue 12, Pages (December 2012)
HyeongJun Kim, Jen Hsin, Yanxin Liu, Paul R. Selvin, Klaus Schulten 
Volume 24, Issue 4, Pages (April 2016)
Structure-Guided Design of Fluorescent S-Adenosylmethionine Analogs for a High- Throughput Screen to Target SAM-I Riboswitch RNAs  Scott F. Hickey, Ming C.
Solution and Crystal Structures of a Sugar Binding Site Mutant of Cyanovirin-N: No Evidence of Domain Swapping  Elena Matei, William Furey, Angela M.
Anindita Dutta, Ivet Bahar  Structure 
Joe G. Greener, Ioannis Filippis, Michael J.E. Sternberg  Structure 
Volume 16, Issue 5, Pages (May 2008)
Zhaoyong Xi, Matthew J. Whitley, Angela M. Gronenborn  Structure 
A Conformational Switch in the CRIB-PDZ Module of Par-6
Volume 23, Issue 11, Pages (November 2015)
Volume 20, Issue 2, Pages (February 2012)
Volume 111, Issue 9, Pages (November 2016)
Discovery Through the Computational Microscope
Structural Analysis of Ligand Stimulation of the Histidine Kinase NarX
Janin Glaenzer, Martin F. Peter, Gavin H. Thomas, Gregor Hagelueken 
Volume 96, Issue 7, Pages (April 2009)
FRET or No FRET: A Quantitative Comparison
Volume 21, Issue 6, Pages (June 2013)
Combining Efficient Conformational Sampling with a Deformable Elastic Network Model Facilitates Structure Refinement at Low Resolution  Gunnar F. Schröder,
Geometry-Based Sampling of Conformational Transitions in Proteins
R.F. Fischetti, D.J. Rodi, D.B. Gore, L. Makowski  Chemistry & Biology 
Volume 110, Issue 12, Pages (June 2016)
Volume 21, Issue 5, Pages (May 2013)
Alemayehu A. Gorfe, Barry J. Grant, J. Andrew McCammon  Structure 
Volume 21, Issue 11, Pages (November 2013)
Saswata Sankar Sarkar, Jayant B. Udgaonkar, Guruswamy Krishnamoorthy 
Volume 23, Issue 9, Pages (September 2015)
Cholesterol Modulates the Dimer Interface of the β2-Adrenergic Receptor via Cholesterol Occupancy Sites  Xavier Prasanna, Amitabha Chattopadhyay, Durba.
The Effect of Dye-Dye Interactions on the Spatial Resolution of Single-Molecule FRET Measurements in Nucleic Acids  Nicolas Di Fiori, Amit Meller  Biophysical.
Volume 13, Issue 7, Pages (July 2005)
Conformational Heterogeneity in the Activation Mechanism of Bax
Tradeoffs and Optimality in the Evolution of Gene Regulation
Atomic-Level Protein Structure Refinement Using Fragment-Guided Molecular Dynamics Conformation Sampling  Jian Zhang, Yu Liang, Yang Zhang  Structure 
Volume 25, Issue 7, Pages e3 (July 2017)
Saswata Sankar Sarkar, Jayant B. Udgaonkar, Guruswamy Krishnamoorthy 
Volume 24, Issue 10, Pages (October 2016)
Volume 20, Issue 1, Pages (January 2012)
Ligand-Driven Vectorial Folding of Ribosome-Bound Human CFTR NBD1
The Atomistic Mechanism of Conformational Transition in Adenylate Kinase: A TEE-REX Molecular Dynamics Study  Marcus B. Kubitzki, Bert L. de Groot  Structure 
Volume 19, Issue 7, Pages (July 2011)
Volume 24, Issue 1, Pages (January 2016)
The Selectivity of K+ Ion Channels: Testing the Hypotheses
Predicting Allosteric Changes from Conformational Ensembles
In Search of the Hair-Cell Gating Spring
Damian Dawidowski, David S. Cafiso  Structure 
Mechanism of Interaction between the General Anesthetic Halothane and a Model Ion Channel Protein, III: Molecular Dynamics Simulation Incorporating a.
Volume 25, Issue 9, Pages e3 (September 2017)
Yongli Zhang, Junyi Jiao, Aleksander A. Rebane  Biophysical Journal 
Po-chia Chen, Jochen S. Hub  Biophysical Journal 
Structural and Thermodynamic Basis for Enhanced DNA Binding by a Promiscuous Mutant EcoRI Endonuclease  Paul J. Sapienza, John M. Rosenberg, Linda Jen-Jacobson 
An Efficient Null Model for Conformational Fluctuations in Proteins
Volume 24, Issue 10, Pages (October 2016)
Presentation transcript:

Volume 21, Issue 1, Pages 9-19 (January 2013) Accurate High-Throughput Structure Mapping and Prediction with Transition Metal Ion FRET  Xiaozhen Yu, Xiongwu Wu, Guillermo A. Bermejo, Bernard R. Brooks, Justin W. Taraska  Structure  Volume 21, Issue 1, Pages 9-19 (January 2013) DOI: 10.1016/j.str.2012.11.013 Copyright © 2013 Elsevier Ltd Terms and Conditions

Structure 2013 21, 9-19DOI: (10.1016/j.str.2012.11.013) Copyright © 2013 Elsevier Ltd Terms and Conditions

Figure 1 Mapping the Structure of MBP with tmFRET (A) The X-ray crystal structures of MBP in the APO (gray, PDB 1OMP) and HOLO (green, PDB 3MBP) states. (B and C) A model of bifunctional labeling of a single protein with a cysteine-linked dye fluoresceine-5-maleimide and a di-histidine coordinated metal ion or (C) cysteine-linked mono-bromobimane and a di-histidine coordinated metal ion. (D) Dynamic distance map comparing the HOLO and APO state crystal structure of MBP. The colormap shows the amino acid to amino acid difference in distance between each state of the protein. The location of the chosen label sites are indicated as a circle (fixed pair) or an x (dynamic pair). (E and F) Cartoon of the label sites in MBP. The pairs that map fixed distances are shown in (E) and pairs that map dynamic distances are shown in (F). Structure 2013 21, 9-19DOI: (10.1016/j.str.2012.11.013) Copyright © 2013 Elsevier Ltd Terms and Conditions

Figure 2 tmFRET Measurements of MBP (A) Cartoon of the experimental system. Fluorescence was measured in a 96-well plate through a fiber-optic launched fluorometer. Increasing concentrations of metal were added to a constant concentration of protein. (B) Spectra were collected from each well and the quantity of quenching was measured. (C) For analysis, spectra were normalized to the initial value at zero metal and the relative fluorescence from a di-histidine containing mutant (solid trace) was plotted as a function of metal concentration. All constructs were compared to cysteine-only controls (dotted lines). From these traces, FRET efficiencies and Kd could be determined. (D) Representative quenching curves for di-histidine-containing MBP mutants (four of ten are shown) compared to cysteine-only controls. Spectra were collected for MBP in the absence (APO, black line) and presence (HOLO, red line) of maltose. Error bars represent standard deviation. See also Figures S1–S5. Structure 2013 21, 9-19DOI: (10.1016/j.str.2012.11.013) Copyright © 2013 Elsevier Ltd Terms and Conditions

Figure 3 Direct Comparison between tmFRET Distances and the X-Ray Crystal Structure Distances (A) Plot of FRET efficiencies between F5M-labeled MBP mutants and nickel compared to the distances in the crystal structures. (B) Plot of distances calculated from FRET between F5M and nickel compared to distances from the X-ray crystal structures. (C) Plot of FRET efficiencies between F5M-labeled MBP mutants and copper compared to the distances in the crystal structures. (D) Plot of distances calculated from FRET between F5M and copper compared to distances from the X-ray crystal structures. (E) Plot of FRET efficiencies between mBBr-labeled MBP mutants and nickel compared to the distances in the crystal structures. (F) Plot of distances calculated from FRET between mBBr and nickel compared to distances from the X-ray crystal structures. (G) Plot of FRET efficiencies between mBBr-labeled MBP mutants and copper compared to the distances in the crystal structures. (H) Plot of distances calculated from FRET between mBBr and copper compared to distances from the X-ray crystal structures. (I) Comparison between the APO state FRET distances and the HOLO state FRET distances for fixed pairs in F5M-labeled MBP donors with nickel (green) or copper (blue) acceptors. (J) The same comparison for dynamic pairs in F5M-labeled MBP. (K) Magnitude of crystal structure calculated distance changes and FRET calculated distance changes for dynamic pairs labeled with F5M. (L) Comparison between the APO state FRET distances and the HOLO state FRET distances for fixed pairs in mBBr-labeled MBP donors with nickel (green) or copper (blue) acceptors. (M) The same comparison for dynamic pairs in mBBr-labeled MBP. (N) Magnitude of crystal structure calculated distance changes and FRET calculated distance changes for dynamic pairs labeled with mBBr. Error bars represent standard deviation. See also Figures S6 and S7. Structure 2013 21, 9-19DOI: (10.1016/j.str.2012.11.013) Copyright © 2013 Elsevier Ltd Terms and Conditions

Figure 4 Using tmFRET Distances to Guide Molecular Simulations SGLD simulations of MBP starting in the HOLO state (PDB 3MBP). Plot compares the RMSD during the simulation between the simulation conformation and the APO state crystal structure of MBP (PDB 1OMP). After 2 ns of simulation, MBP does not find the APO state (black trace). With tmFRET-derived distance constraints added to the simulation, the simulation adopts the APO state within 2 ns (gray trace). Representative structures from the tmFRET-constrained simulation are shown below. The color of the simulated structures matches time-points on the graph. As a comparison, the gray structure is the APO state crystal structure. See also Figures S8 and S9 and Movies S1 and S2. Structure 2013 21, 9-19DOI: (10.1016/j.str.2012.11.013) Copyright © 2013 Elsevier Ltd Terms and Conditions