Transmission electron microscopy as a tool for nanocrystal characterization pre- and post-injector.
Philos Trans R Soc Lond B Biol Sci
; 369(1647): 20130322, 2014 Jul 17.
Article
de En
| MEDLINE
| ID: mdl-24914151
ABSTRACT
Recent advancements at the Linac Coherent Light Source X-ray free-electron laser (XFEL) enabling successful serial femtosecond diffraction experiments using nanometre-sized crystals (NCs) have opened up the possibility of X-ray structure determination of proteins that produce only submicrometre crystals such as many membrane proteins. Careful crystal pre-characterization including compatibility testing of the sample delivery method is essential to ensure efficient use of the limited beamtime available at XFEL sources. This work demonstrates the utility of transmission electron microscopy for detecting and evaluating NCs within the carrier solutions of liquid injectors. The diffraction quality of these crystals may be assessed by examining the crystal lattice and by calculating the fast Fourier transform of the image. Injector reservoir solutions, as well as solutions collected post-injection, were evaluated for three types of protein NCs (i) the membrane protein PTHR1, (ii) the multi-protein complex Pol II-GFP and (iii) the soluble protein lysozyme. Our results indicate that the concentration and diffraction quality of NCs, particularly those with high solvent content and sensitivity to mechanical manipulation may be affected by the delivery process.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Diffraction des rayons X
/
Microscopie électronique à transmission
/
Électrons
/
Nanoparticules
/
Lasers
/
Conformation moléculaire
Limites:
Humans
Langue:
En
Journal:
Philos Trans R Soc Lond B Biol Sci
Année:
2014
Type de document:
Article
Pays d'affiliation:
États-Unis d'Amérique