Journal:IUCrJ:S2052252525006645
From Proteopedia
(Difference between revisions)

(45 intermediate revisions not shown.) | |||
Line 1: | Line 1: | ||
- | <StructureSection load='' size='450' side='right' scene=' | + | <StructureSection load='' size='450' side='right' scene='10/1087716/020_Fig_S8a/1' caption=''> |
===Time-resolved serial synchrotron and serial femtosecond crystallography of heme proteins using photocaged nitric oxide=== | ===Time-resolved serial synchrotron and serial femtosecond crystallography of heme proteins using photocaged nitric oxide=== | ||
- | <big>Peter Smyth, Sofia Jaho, Lewis J. Williams, | + | <big>Peter Smyth, Sofia Jaho, Lewis J. Williams, Gabriel Karras, Ann Fitzpatrick, Amy J. Thompson,a Sinan Battah, Danny Axford, Sam Horrell, Marina Lučić, Kotone Ishihara, Machika Kataoka, Hiroaki Matsuura, Kanji Shimba, Kensuke Tono, Takehiko Tosha, Hiroshi Sugimoto, Shigeki Owada, Michael A. Hough, Jonathan A.R. Worrall, Robin L. Owen</big> <ref>doi: 10.1107/S2052252525006645</ref> |
<hr/> | <hr/> | ||
<b>Molecular Tour</b><br> | <b>Molecular Tour</b><br> | ||
Line 9: | Line 9: | ||
Laser-activated SSX structures of DtpB. Omit electron density, contoured at 5σ maps for NO-bound heme site in chain C of DtpB, collected with different laser energies of <br> | Laser-activated SSX structures of DtpB. Omit electron density, contoured at 5σ maps for NO-bound heme site in chain C of DtpB, collected with different laser energies of <br> | ||
- | <scene name='10/1087716/ | + | <scene name='10/1087716/020_Fig_6a_lab_sp_pse/1'>.81</scene> '''μJ''' <br> |
- | 8.1 μJ<br> | + | <scene name='10/1087716/020_Fig_6b_lab_sp_pse/1'>8.1</scene> '''μJ''' <br> |
- | 16.1 μJ<br> | + | <scene name='10/1087716/020_Fig_6c_lab_sp_spe/1'>16.1</scene> '''μJ''' <br> |
- | 32.2 μJ<br> | + | <scene name='10/1087716/020_Fig_6d_lab_sp_pse/1'>32.2</scene> '''μJ''' <br> |
- | 64.4 μJ<br> | + | <scene name='10/1087716/020_Fig_6e_lab_sp_pse/1'>64.4</scene> '''μJ''' <br> |
- | + | <br> | |
+ | <jmol> | ||
+ | <jmolButton> | ||
+ | <text>animation</text> | ||
+ | <script> | ||
+ | var a = [1,2,3,4]; | ||
+ | for(var i IN a) { | ||
+ | script /wiki/scripts/10/1087716/020_Fig_6a_lab_sp_pse/1.spt; delay 0.8; | ||
+ | script /wiki/scripts/10/1087716/020_Fig_6b_lab_sp_pse/1.spt; delay 0.8; | ||
+ | script /wiki/scripts/10/1087716/020_Fig_6c_lab_sp_spe/1.spt; delay 0.8; | ||
+ | script /wiki/scripts/10/1087716/020_Fig_6d_lab_sp_pse/1.spt; delay 0.8; | ||
+ | script /wiki/scripts/10/1087716/020_Fig_6e_lab_sp_pse/1.spt; delay 0.8; | ||
+ | } | ||
+ | </script> | ||
+ | </jmolButton> | ||
+ | </jmol> | ||
+ | With increasing laser power, the electron density maps support the presence of an NO molecule bound to the heme iron with increasing partial occupancy. | ||
- | Laser parameters for photoactivation are systematically explored, and time-resolved structures over timescales ranging from 100 µs to 1.4 s using synchrotron and XFEL beamlines are described. The effective use of this photocage for time-resolved crystallography is demonstrated with laser powers as low as 0.19 µJ (~0.1 nJ∙µm | + | Laser parameters for photoactivation are systematically explored, and time-resolved structures over timescales ranging from 100 µs to 1.4 s using synchrotron and XFEL beamlines are described. The effective use of this photocage for time-resolved crystallography is demonstrated with laser powers as low as 0.19 µJ (~0.1 nJ∙µm<sup>-2</sup>), resulting in high occupancy NO-bound protein structures. Appropriate illumination conditions for such experiments are determined, and considerations for more general reaction initiation using photocages are discussed. An example is seen in the results of a high laser intensity illumination (1600 mJ, PDB: [[9htc]]) McCP-β showing maps and chain A, for a <scene name='10/1087716/020_Fig_S8a/1'>2Fo minus Fc</scene> and for an <scene name='10/1087716/020_Fig_S8b/1'>omit</scene>, contoured at 1σ and 5σ respectively. In this experiment, the fixed target chip was reversed in orientation compared to the other structures in this manuscript, and Mylar film was used to seal the chip, with the result that far less light reached the sample. |
<b>References</b><br> | <b>References</b><br> |
Current revision
|
This page complements a publication in scientific journals and is one of the Proteopedia's Interactive 3D Complement pages. For aditional details please see I3DC.