Watching proteins move using site-directed spin labeling

Slides:



Advertisements
Similar presentations
Volume 28, Issue 4, Pages (November 2007)
Advertisements

Volume 127, Issue 4, Pages (November 2006)
Munirathinam Sundaramoorthy, James Terner, Thomas L Poulos  Structure 
Volume 26, Issue 1, Pages (April 2007)
Investigation of RNA-Protein and RNA-Metal Ion Interactions by Electron Paramagnetic Resonance Spectroscopy  Thomas E Edwards, Tamara M Okonogi, Snorri.
Volume 9, Issue 10, Pages (October 2001)
The loop E–loop D region of Escherichia coli 5S rRNA: the solution structure reveals an unusual loop that may be important for binding ribosomal proteins 
Volume 13, Issue 6, Pages (March 2004)
Structural Basis for the Highly Selective Inhibition of MMP-13
Structural States and Dynamics of the D-Loop in Actin
Conformational changes in rhodopsin Example Lecture
Volume 10, Issue 7, Pages (July 2002)
Volume 23, Issue 9, Pages (September 2015)
Crystal Structure of Chicken γS-Crystallin Reveals Lattice Contacts with Implications for Function in the Lens and the Evolution of the βγ-Crystallins 
Electron Paramagnetic Resonance
A Distance Ruler for RNA Using EPR and Site-Directed Spin Labeling
Conformational Change in an MFS Protein: MD Simulations of LacY
The TNF and TNF Receptor Superfamilies
Volume 24, Issue 7, Pages (July 2016)
Structure of an LDLR-RAP Complex Reveals a General Mode for Ligand Recognition by Lipoprotein Receptors  Carl Fisher, Natalia Beglova, Stephen C. Blacklow 
Daniel M. Freed, Peter S. Horanyi, Michael C. Wiener, David S. Cafiso 
Volume 6, Issue 10, Pages (October 1998)
Volume 11, Issue 11, Pages (November 2003)
Tamas Yelland, Snezana Djordjevic  Structure 
Volume 12, Issue 12, Pages (December 2004)
UG Wagner, M Hasslacher, H Griengl, H Schwab, C Kratky  Structure 
Volume 106, Issue 6, Pages (March 2014)
Volume 28, Issue 4, Pages (November 2007)
Volume 108, Issue 6, Pages (March 2015)
A Model for the Solution Structure of the Rod Arrestin Tetramer
Microsecond Unfolding Kinetics of Sheep Prion Protein Reveals an Intermediate that Correlates with Susceptibility to Classical Scrapie  Kai-Chun Chen,
Einav Gross, David B Kastner, Chris A Kaiser, Deborah Fass  Cell 
Volume 17, Issue 8, Pages (August 2010)
Volume 19, Issue 12, Pages (December 2011)
Christian Kappel, Ulrich Zachariae, Nicole Dölker, Helmut Grubmüller 
De Novo Design of Foldable Proteins with Smooth Folding Funnel
Volume 31, Issue 2, Pages (July 2008)
Beena Krishnan, Lila M. Gierasch  Chemistry & Biology 
Volume 21, Issue 10, Pages (October 2013)
Munirathinam Sundaramoorthy, James Terner, Thomas L Poulos  Structure 
Volume 111, Issue 9, Pages (November 2016)
Volume 124, Issue 5, Pages (March 2006)
Volume 9, Issue 8, Pages (August 2001)
Volume 13, Issue 8, Pages (August 2005)
Daniel Peisach, Patricia Gee, Claudia Kent, Zhaohui Xu  Structure 
Volume 26, Issue 1, Pages (April 2007)
Anne Dallas, Harry F Noller  Molecular Cell 
Structural Basis for the Highly Selective Inhibition of MMP-13
A Different Look for AB5 Toxins
Volume 6, Issue 1, Pages (July 2000)
Volume 23, Issue 9, Pages (September 2015)
Saswata Sankar Sarkar, Jayant B. Udgaonkar, Guruswamy Krishnamoorthy 
Volume 110, Issue 7, Pages (April 2016)
Volume 114, Issue 3, Pages (February 2018)
Allosteric Control of Syntaxin 1a by Munc18-1: Characterization of the Open and Closed Conformations of Syntaxin  Damian Dawidowski, David S. Cafiso 
Miyeon Kim, Qi Xu, Gail E. Fanucci, David S. Cafiso 
Volume 4, Issue 2, Pages (February 1996)
Structure of a water soluble fragment of the ‘Rieske’ iron–sulfur protein of the bovine heart mitochondrial cytochrome bc1 complex determined by MAD phasing.
Volume 6, Issue 2, Pages (August 2000)
Jia-Wei Wu, Amy E. Cocina, Jijie Chai, Bruce A. Hay, Yigong Shi 
Volume 10, Issue 5, Pages (May 2002)
Damian Dawidowski, David S. Cafiso  Structure 
Volume 25, Issue 9, Pages e3 (September 2017)
A Model for the Solution Structure of the Rod Arrestin Tetramer
Volume 27, Issue 1, Pages (July 2007)
Christian Kappel, Ulrich Zachariae, Nicole Dölker, Helmut Grubmüller 
Dynamic Short Hydrogen Bonds in Histidine Tetrad of Full-Length M2 Proton Channel Reveal Tetrameric Structural Heterogeneity and Functional Mechanism 
Yogesh K. Gupta, Deepak T. Nair, Robin P. Wharton, Aneel K. Aggarwal 
Volume 9, Issue 2, Pages (February 2001)
Volume 13, Issue 6, Pages (March 2004)
Presentation transcript:

Watching proteins move using site-directed spin labeling Wayne L Hubbell, Hassane S Mchaourab, Christian Altenbach, Michael A Lietzow  Structure  Volume 4, Issue 7, Pages 779-783 (July 1996) DOI: 10.1016/S0969-2126(96)00085-8

Figure 1 Reaction of the methanethiosulfonate spin label I with cysteine to generate the disulfide-linked nitroxide side chain R1. Structure 1996 4, 779-783DOI: (10.1016/S0969-2126(96)00085-8)

Figure 2 Determination of secondary structure with nitroxide scanning. The upper panel shows structures of T4L and CRBP with sites of single nitroxide substitutions indicated by magenta spheres. The lower panel shows plots of II(O2) (red) and ΔHo−1 (yellow) as a function of sequence number for the nitroxide scans. The smooth trace in the data for T4L is a function with a period of 3.6. Structure 1996 4, 779-783DOI: (10.1016/S0969-2126(96)00085-8)

Figure 3 The inverse spectral second moments (<H2>−1) and the inverse central linewidths (ΔHo−1) for the R1 side chain at twenty sites in T4L. The classifications according to protein topography are discussed in the text. (Figure was adapted from [5], with permission.) Structure 1996 4, 779-783DOI: (10.1016/S0969-2126(96)00085-8)

Figure 4 The interdomain helix in T4L, showing an engineered binding site for Cu(II) consisting of two histidine residues (His65 and His69), and a nitroxide side chain (R1, at residue 80). Distance (r) from the copper site to single R1 side chains, determined from magnetic interactions, is consistent with those estimated from molecular modeling [19]. Structure 1996 4, 779-783DOI: (10.1016/S0969-2126(96)00085-8)

Figure 5 Time-resolved folding in T4L. (a) T4L structure showing the locations of Ile3→R1and Val71→R1. In the folded structure, the nitroxide residues are predicted to be essentially in van der Waals contact. (b) The equilibrium EPR spectra of the doubly labeled T4L in the native state at pH 6.8 (red traces) and unfolded state in the presence of 3M urea at pH 2.0 (yellow trace). The lower spectrum is for the native protein with a 30x increase in gain to reveal the lineshape of the interacting spins. (c) Time-resolved intensity changes recorded at the position of the arrow in (b) following rapid mixing of the unfolded protein with a buffer to produce a jump in pH from 2.0 to 6.8 to initiate the folding reaction. Structure 1996 4, 779-783DOI: (10.1016/S0969-2126(96)00085-8)