Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

Structural Dynamics
E Han DaoHasan Demirci

Abstract

In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS) using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecond X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a com...Continue Reading

References

Jan 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·M WeikJ L Sussman
Apr 4, 2000·Structure·R B Ravelli, S M McSweeney
Oct 3, 2000·Nature·B T WimberlyV Ramakrishnan
Jul 13, 2004·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Stefan P Hau-RiegeAbraham Szoke
Nov 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·A Gonzalez, C Nave
Dec 17, 2004·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Magnus BerghDavid van der Spoel
Mar 22, 2006·Proceedings of the National Academy of Sciences of the United States of America·Robin Leslie OwenElspeth F Garman
Jun 1, 2004·Journal of Applied Crystallography·Nicholas K SauterPaul D Adams
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Wolfgang Kabsch
Dec 31, 2010·Biophysical Journal·M S HunterP Fromme
Feb 5, 2011·Nature·Henry N ChapmanJohn C H Spence
Jun 6, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jan KernVittal K Yachandra
Oct 24, 2012·Acta Crystallographica. Section D, Biological Crystallography·Raymond G SierraMichael J Bogan
Jan 17, 2013·Nature Communications·Hasan DemirciGerwald Jogl
Jun 26, 2013·Acta Crystallographica. Section D, Biological Crystallography·Nicholas K SauterNathaniel Echols
Aug 31, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Hasan DemirciMichael J Bogan
Jan 1, 2012·Nature Photonics·Anton BartyHenry N Chapman
Dec 1, 2011·Physical Review. B, Condensed Matter and Materials Physics·Lukas LombIlme Schlichting
Dec 21, 2013·Science·Wei LiuVadim Cherezov
Feb 15, 2014·Nature Communications·Uwe WeierstallVadim Cherezov
Jun 7, 2014·Journal of Applied Crystallography·Anton BartyHenry Chapman
Jun 28, 2014·Nature Communications·B F MurphyN Berrah
Nov 2, 2014·Proceedings of the National Academy of Sciences of the United States of America·Aina E CohenKeith O Hodgson
Feb 11, 2015·Acta Crystallographica. Section D, Biological Crystallography·Oliver B ZeldinAxel T Brunger
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

❮ Previous
Next ❯

Methods Mentioned

BETA
X-ray

Software Mentioned

Web
Ice
SPOTFINDER
HKL
LABELIT
Blu
ProtParam
XFEL

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
A E CohenK O Hodgson
Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Hasan DemirciMichael J Bogan
Physical Review. B, Condensed Matter and Materials Physics
Lukas LombIlme Schlichting
© 2022 Meta ULC. All rights reserved