A5051385736- Enzyme Structure and Function
- Photosynthetic Processes and Mechanisms
- Photoreceptor and optogenetics research
- Light effects on plants
- Protein Structure and Dynamics
- Hemoglobin structure and function
- Advanced X-ray Imaging Techniques
- Mass Spectrometry Techniques and Applications
- bioluminescence and chemiluminescence research
- Heme Oxygenase-1 and Carbon Monoxide
- Neonatal Health and Biochemistry
- X-ray Spectroscopy and Fluorescence Analysis
- Erythrocyte Function and Pathophysiology
- Chronic Disease Management Strategies
- Advanced Electron Microscopy Techniques and Applications
- Biocrusts and Microbial Ecology
- Retinal Development and Disorders
- Crystallography and Radiation Phenomena
- X-ray Diffraction in Crystallography
- Spectroscopy and Quantum Chemical Studies
- Microbial Community Ecology and Physiology
- RNA and protein synthesis mechanisms
- Advanced Fluorescence Microscopy Techniques
- Algal biology and biofuel production
- Hemoglobinopathies and Related Disorders
Public Health Scotland
2021-2024
University of St Andrews
2019-2023
University of Chicago
2012-2022
University of Glasgow
2008-2020
University of Illinois Chicago
1995-2016
Argonne National Laboratory
2003-2012
Cancer Research UK Scotland Institute
2008
University of Washington
2008
Molecular Biology Consortium
1996-2007
Tokyo Institute of Technology
2005-2006
The biological activity of macromolecules is accompanied by rapid structural changes. photosensitivity the carbon monoxide complex myoglobin was used at European Synchrotron Radiation Facility to obtain pulsed, Laue x-ray diffraction data with nanosecond time resolution during process heme and protein relaxation after photodissociation rebinding. These time-resolved experiments reveal structures photoproducts, provide a foundation spectroscopic results molecular dynamics calculations,...
Serial femtosecond crystallography using ultrashort pulses from x-ray free electron lasers (XFELs) enables studies of the light-triggered dynamics biomolecules. We used microcrystals photoactive yellow protein (a bacterial blue light photoreceptor) as a model system and obtained high-resolution, time-resolved difference density maps excellent quality with strong features; these allowed determination structures reaction intermediates to resolution 1.6 angstroms. Our results open way study...
Phototropin, a major blue-light receptor for phototropism in seed plants, exhibits blue-light-dependent autophosphorylation and contains two light, oxygen, or voltage (LOV) domains serine/threonine kinase domain. The LOV share homology with the PER-ARNT-SIM (PAS) superfamily, diverse group of sensor proteins. Each domain noncovalently binds single FMN molecule reversible photochemistry vitro when expressed separately tandem. We have determined crystal structure LOV2 from phototropin segment...
Visualizing a response to light Many biological processes depend on detecting and responding light. The is often mediated by structural change in protein that begins when absorption of photon causes isomerization chromophore bound the protein. Pande et al. used x-ray pulses emitted free electron laser source conduct time-resolved serial femtosecond crystallography time range 100 fs 3 ms. This allowed for real-time tracking trans-cis photoactive yellow associated changes Science , this issue p. 725
For single-cell and multicellular systems to survive, they must accurately sense respond their cellular extracellular environment. Light is a nearly ubiquitous environmental factor, many species have evolved the capability this stimulus. Numerous photoreceptors underlie activation of light-sensitive signal transduction cascades controlling these responses. Here, we review properties light, oxygen, or voltage (LOV) family blue-light photoreceptor domains, subset Per-ARNT-Sim (PAS)...
The blue-light photoreceptor photoactive yellow protein (PYP) undergoes a self-contained light cycle. atomic structure of the bleached signaling intermediate in cycle PYP was determined by millisecond time-resolved, multiwavelength Laue crystallography and simultaneous optical spectroscopy. Light-induced trans-to-cis isomerization 4-hydroxycinnamyl chromophore coupled rearrangements produce new set active-site hydrogen bonds. An arginine gateway opens, allowing solvent exposure protonation...
The structure of bovine intestinal calcium-binding protein (ICaBP) has been determined crystallographically at a resolution 2.3 A and refined by least
The management of people with multiple chronic conditions challenges health-care systems designed around single conditions. There is international consensus that care for multimorbidity should be patient-centred, focus on quality life, and promote self-management towards agreed goals. However, there little evidence about the effectiveness this approach. Our hypothesis was so-called 3D approach (based dimensions health, depression, drugs) patients would improve their health-related which...
Light sensing by photoreceptors controls phototropism, chloroplast movement, stomatal opening, and leaf expansion in plants. Understanding the molecular mechanism which these processes are regulated requires a quantitative description of photoreceptor dynamics. We focus on light-driven signal transduction LOV2 domain (LOV, light, oxygen, voltage) blue light phototropin 1 from Avena sativa (oat). High-resolution crystal structures dark states an oat construct including residues Leu404 through...
An understanding of how allostery, the conformational coupling distant functional sites, arises in highly evolvable systems is considerable interest areas ranging from cell biology to protein design and signaling networks. We reasoned that rigidity defined geometry an alpha-helical domain linker would make it effective as a conduit for allosteric signals. To test this idea, we rationally designed 12 fusions between naturally photoactive LOV2 Avena sativa phototropin 1 Escherichia coli trp...
Phytochromes are red-light photoreceptors that regulate light responses in plants, fungi, and bacteria via reversible photoconversion between red (Pr) far-red (Pfr) light-absorbing states. Here we report the crystal structure at 2.9 A resolution of a bacteriophytochrome from Pseudomonas aeruginosa with an intact, fully photoactive photosensory core domain its dark-adapted Pfr state. This reveals how unusual interdomain interactions, including knot "arm" near chromophore site, bring together...
Multimorbidity is a growing issue and poses major challenge to health care systems around the world. related ageing but many studies have now shown that it also socially patterned, being more common occurring at an earlier age in areas of high socioeconomic deprivation. There lack research on patients with multimorbidity, thus guidelines are based single-conditions. Polypharmacy increasing drug-disease drug-drug interactions. Multimorbid need holistic care, secondary services highly...
The phototropins are flavoprotein kinases that control phototropic bending, light-induced chloroplast movement, and stomatal opening in plants. Two flavin mononucleotide binding light, oxygen, or voltage (LOV) domains the sites for initial photochemistry these blue light photoreceptors. We have determined steady state, photoexcited crystal structure of a flavin-bound LOV domain. reveals unique photochemical switch pocket which absorption drives formation reversible covalent bond between...
A time-resolved Laue X-ray diffraction technique has been used to explore protein relaxation and ligand migration at room temperature following photolysis of a single crystal carbon monoxymyoglobin. The CO is photodissociated by 7.5 ns laser pulse, the subsequent structural changes are probed 150 ps or 1 μs pulses 14 laser/X-ray delay times, ranging from 1.9 ms. Very fast heme involving E F helices evident data time delay. molecules detected two locations: distal pocket docking site Xe...
Photoactive yellow protein (PYP) is a member of the xanthopsin family eubacterial blue-light photoreceptors. On absorption light, PYP enters photocycle that ultimately transduces energy contained in light signal into an altered biological response. Nanosecond time-resolved x-ray crystallography was used to determine structure short-lived, red-shifted, intermediate state denoted [pR], which develops within 1 nanosecond after photoelectronic excitation chromophore by light. The resulting...
Determining 3D intermediate structures during the biological action of proteins in real time under ambient conditions is essential for understanding how function. Here we use time-resolved Laue crystallography to extract short-lived and thereby unveil signal transduction blue light photoreceptor photoactive yellow protein (PYP) from Halorhodospira halophila. By analyzing a comprehensive set data PYP photocycle (forty-seven points one nanosecond second), track all atoms its directly observe...
The flavin-binding BLUF domain of AppA represents a new class blue light photoreceptors that are present in number bacterial and algal species. dark state X-ray structure this was determined at 2.3 Å resolution. demonstrates function for the common ferredoxin-like fold; two long α-helices flank flavin, which is bound with its isoalloxazine ring perpendicular to five-stranded β-sheet. hydrogen bond network overall protein topology (but not sequence) bear some resemblance LOV domains, subset...
The phototropins constitute an important class of plant photoreceptor kinases that control a range physiological responses, including phototropism, light-directed chloroplast movement, and light-induced stomatal opening. LOV2 domain phototropin binds molecule flavin mononucleotide (FMN) undergoes photocycle involving light-driven covalent adduct formation between conserved cysteine residue the C(4a) atom FMN. This product state promotes C-terminal kinase activation downstream signal...