Imaging and structural studies of DNA-protein complexes and membrane ion channels.
In bio-imaging by electron microscopy, damage of the sample and limited contrast are the two main hurdles for reaching high image quality. We extend a new preparation method based on nanofabrication and super-hydrophobicity to the imaging and structural studies of nucleic acids, nucleic acid-protein complexes (DNA/Rad51 repair protein complex) and neuronal ion channels (gap-junction, K+and GABAAchannels) as paradigms of biological significance and increasing complexity. The preparation method is based on the liquid phase and is compatible with physiological conditions. Only in the very last stage, samples are dried for TEM analysis. Conventional TEM and high-resolution TEM (HRTEM) were used to achieve a resolution of 3.3 and 1.5 Å, respectively. The EM dataset quality allows the determination of relevant structural and metrological information on the DNA structure, DNA-protein interactions and ion channels, allowing the identification of specific macromolecules and their structure.
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
M
M
Marini
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
T
T
Limongi
King Abdullah University of Science and Technology, NABLA lab, BESE Division, Thuwal, 23955-6900, Kingdom of Saudi Arabia.
A
http://orcid.org/0000-0002-1476-7742
A
Falqui
King Abdullah University of Science and Technology, NABLA lab, BESE Division, Thuwal, 23955-6900, Kingdom of Saudi Arabia.
A
A
Genovese
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
M
M
Allione
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
M
M
Moretti
King Abdullah University of Science and Technology, Imaging and Characterization core lab, Thuwal, 23955-6900, Kingdom of Saudi Arabia.
S
S
Lopatin
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
L
L
Tirinato
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
G
G
Das
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
B
B
Torre
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
A
A
Giugni
Center for Synaptic Neuroscience and Technology, Istituto Italiano di Tecnologia, Genova, Italy.
F
F
Cesca
Center for Synaptic Neuroscience and Technology, Istituto Italiano di Tecnologia, Genova, Italy.
F
F
Benfenati
King Abdullah University of Science and Technology, SMILEs lab, PSE and BESE Divisions, Thuwal, 23955-6900, Kingdom of Saudi Arabia. enzo.difabrizio@kaust.edu.sa.
E
http://orcid.org/0000-0001-5886-4678
E
Di Fabrizio
D000818:Animals; D004247:DNA / Q000648:ultrastructure*; D007473:Ion Channels / Q000648:ultrastructure*; D008051:Lipid Bilayers; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D009474:Neurons / Q000648:ultrastructure; D009696:Nucleic Acids / Q000648:ultrastructure*; D051135:Rad51 Recombinase / Q000648:ultrastructure
D007473:Ion Channels; D008051:Lipid Bilayers; D009696:Nucleic Acids; D004247:DNA; D051135:Rad51 Recombinase