UV laser-excited fluorescence as a tool for the visualization of protein crystals mounted in loops

Acta Crystallographica. Section D, Biological Crystallography
Xavier VernedeDominique Bourgeois

Abstract

Structural proteomics has promoted the rapid development of automated protein structure determination using X-ray crystallography. Robotics are now routinely used along the pipeline from genes to protein structures. However, a bottleneck still remains. At synchrotron beamlines, the success rate of automated sample alignment along the X-ray beam is limited by difficulties in visualization of protein crystals, especially when they are small and embedded in mother liquor. Despite considerable improvement in optical microscopes, the use of visible light transmitted or reflected by the sample may result in poor or misleading contrast. Here, the endogenous fluorescence from aromatic amino acids has been used to identify even tiny or weakly fluorescent crystals with a high success rate. The use of a compact laser at 266 nm in combination with non-fluorescent sample holders provides an efficient solution to collect high-contrast fluorescence images in a few milliseconds and using standard camera optics. The best image quality was obtained with direct illumination through a viewing system coaxial with the UV beam. Crystallographic data suggest that the employed UV exposures do not generate detectable structural damage.

Citations

Jun 8, 2010·Journal of Structural and Functional Genomics·M CymborowskiW Minor
Sep 21, 2011·Acta Crystallographica. Section D, Biological Crystallography·Jeremy T MaddenGarth J Simpson
Jan 1, 2013·Acta Crystallographica. Section D, Biological Crystallography·Emma L DeWaltGarth J Simpson
Feb 26, 2009·Journal of Synchrotron Radiation·John McGeehanElspeth F Garman
Apr 29, 2011·Journal of Synchrotron Radiation·Daniele de SanctisSantosh Panjikar
Jun 21, 2011·Journal of Synchrotron Radiation·Anuschka PauluhnClemens Schulze-Briese
Jun 15, 2013·Journal of Synchrotron Radiation·Jeremy T MaddenGarth J Simpson
Mar 9, 2010·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Harindarpal S Gill
May 2, 2013·Acta Crystallographica. Section D, Biological Crystallography·David J KissickGarth J Simpson
Mar 5, 2016·Journal of Applied Crystallography·Tiit LukkWarren R Zipfel
Mar 10, 2016·Acta Crystallographica. Section D, Structural Biology·Daniele de SanctisMax H Nanao
Aug 10, 2010·Acta Crystallographica. Section D, Biological Crystallography·David WattsMatthew R Groves
Sep 22, 2012·Journal of Applied Crystallography·I KhanU Englich
Aug 3, 2011·Journal of Applied Crystallography·Sergey StepanovRobert F Fischetti
Apr 6, 2016·Archives of Biochemistry and Biophysics·Robin L OwenMartin Fuchs
Jul 7, 2015·Acta Crystallographica. Section D, Biological Crystallography·Justin A NewmanGarth J Simpson
Jan 24, 2015·Acta Crystallographica. Section D, Biological Crystallography·David von StettenAntoine Royant
Jul 1, 2016·Journal of Synchrotron Radiation·Justin A NewmanGarth J Simpson
Dec 24, 2016·Journal of Synchrotron Radiation·Nicole M ScarboroughGarth J Simpson
Apr 14, 2018·Acta Crystallographica. Section D, Structural Biology·Igor MelnikovAlexander Popov
Jul 6, 2019·Journal of Synchrotron Radiation·Sho ItoMasaki Yamamoto
Apr 8, 2021·Acta Crystallographica. Section D, Structural Biology·Chen LiGarth Simpson
Nov 25, 2020·The Journal of Physical Chemistry. B·Antara VaidyanathanBrahmananda Chakraborty

❮ Previous
Next ❯

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.