Matching Items (61)
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Description
Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the

Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
ContributorsEdlund, Petra (Author) / Takala, Heikki (Author) / Claesson, Elin (Author) / Henry, Leocadie (Author) / Dods, Robert (Author) / Lehtivuori, Heli (Author) / Panman, Matthijs (Author) / Pande, Kanupriya (Author) / White, Thomas (Author) / Nakane, Takanori (Author) / Berntsson, Oskar (Author) / Gustavsson, Emil (Author) / Bath, Petra (Author) / Modi, Vaibhav (Author) / Roy Chowdhury, Shatabdi (Author) / Zook, James (Author) / Berntsen, Peter (Author) / Pandey, Suraj (Author) / Poudyal, Ishwor (Author) / Tenboer, Jason (Author) / Kupitz, Christopher (Author) / Barty, Anton (Author) / Fromme, Petra (Author) / Koralek, Jake D. (Author) / Tanaka, Tomoyuki (Author) / Spence, John (Author) / Liang, Mengning (Author) / Hunter, Mark S. (Author) / Boutet, Sebastien (Author) / Nango, Eriko (Author) / Moffat, Keith (Author) / Groenhof, Gerrit (Author) / Ihalainen, Janne (Author) / Stojkovic, Emina A. (Author) / Schmidt, Marius (Author) / Westenhoff, Sebastian (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2016-10-19
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Description
Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement.

Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1 mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.
ContributorsNogly, Przemyslaw (Author) / Panneels, Valerie (Author) / Nelson, Garrett (Author) / Gati, Cornelius (Author) / Kimura, Tetsunari (Author) / Milne, Christopher (Author) / Milathianaki, Despina (Author) / Kubo, Minoru (Author) / Wu, Wenting (Author) / Conrad, Chelsie (Author) / Coe, Jesse (Author) / Bean, Richard (Author) / Zhao, Yun (Author) / Bath, Petra (Author) / Dods, Robert (Author) / Harimoorthy, Rajiv (Author) / Beyerlein, Kenneth R. (Author) / Rheinberger, Jan (Author) / James, Daniel (Author) / Deponte, Daniel (Author) / Li, Chufeng (Author) / Sala, Leonardo (Author) / Williams, Garth J. (Author) / Hunter, Mark S. (Author) / Koglin, Jason E. (Author) / Berntsen, Peter (Author) / Nango, Eriko (Author) / Iwata, So (Author) / Chapman, Henry N. (Author) / Fromme, Petra (Author) / Frank, Matthias (Author) / Abela, Rafael (Author) / Boutet, Sebastien (Author) / Barty, Anton (Author) / White, Thomas A. (Author) / Weierstall, Uwe (Author) / Spence, John (Author) / Neutze, Richard (Author) / Schertler, Gebhard (Author) / Standfuss, Jorg (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / School of Molecular Sciences (Contributor)
Created2016-08-22
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Description
Because collective cognition emerges from local signaling among group members, deciphering communication systems is crucial to understanding the underlying mechanisms. Alarm signals are widespread in the social insects and can elicit a variety of behavioral responses to danger, but the functional plasticity of these signals has not been well studied.

Because collective cognition emerges from local signaling among group members, deciphering communication systems is crucial to understanding the underlying mechanisms. Alarm signals are widespread in the social insects and can elicit a variety of behavioral responses to danger, but the functional plasticity of these signals has not been well studied. Here we report an alarm pheromone in the ant Temnothorax rugatulus that elicits two different behaviors depending on context. When an ant was tethered inside an unfamiliar nest site and unable to move freely, she released a pheromone from her mandibular gland that signaled other ants to reject this nest as a potential new home, presumably to avoid potential danger. When the same pheromone was presented near the ants' home nest, they were instead attracted to it, presumably to respond to a threat to the colony. We used coupled gas chromatography/mass spectrometry to identify candidate compounds from the mandibular gland and tested each one in a nest choice bioassay. We found that 2,5-dimethylpyrazine was sufficient to induce rejection of a marked new nest and also to attract ants when released at the home nest. This is the first detailed investigation of chemical communication in the leptothoracine ants. We discuss the possibility that this pheromone's deterrent function can improve an emigrating colony's nest site selection performance.
Created2014-09-01
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Description
The Energiewende aims to drastically reduce Germany’s greenhouse gas emissions, without relying on nuclear power, while maintaining a secure and affordable energy supply. Since 2000 the country’s renewable-energy share has increased exponentially, accounting in 2017 for over a third of Germany's gross electricity consumption. This unprecedented achievement is the result

The Energiewende aims to drastically reduce Germany’s greenhouse gas emissions, without relying on nuclear power, while maintaining a secure and affordable energy supply. Since 2000 the country’s renewable-energy share has increased exponentially, accounting in 2017 for over a third of Germany's gross electricity consumption. This unprecedented achievement is the result of policies, tools, and institutional arrangements intended to steer society to a low-carbon economy. Despite its resounding success in renewable-energy deployment, the Energiewende is not on track to meet its decarbonization goals. Energiewende rules and regulations have generated numerous undesired consequences, and have cost much more than anticipated, a burden borne primarily by energy consumers. Why has the Energiewende not only made energy more expensive, but also failed to bring Germany closer to its decarbonization goals? I analyzed the Energiewende as a complex socio-technical system, examining its legal framework and analyzing the consequences of successive regulations; identifying major political and energy players and the factors that motivated them to pursue socio-technical change; and documenting the political trends and events in which the Energiewende is rooted and which continue to shape it. I analyzed the dynamics and the loopholes that created barriers to transition, pushed the utility sector to the brink of dissolution, and led to such undesirable outcomes as negative wholesale prices and forced exports of electricity to Germany’s European neighbors. Thirty high-level energy experts and stakeholders were interviewed to find out how the best-informed members of German society perceive the Energiewende. Surprisingly, although they were highly critical of the way the transition has unfolded, most were convinced that the transition would eventually succeed. But their definitions of success did not always depend on achieving carbon-mitigation targets. Indeed, Germany jeopardizes the achievement of these targets by changing too many policy and institutional variables at too fast a pace. Good intentions and commitment are not enough to create economies based on intermittent energy sources: they will also require intensive grid expansion and breakthroughs in storage technology. The Energiewende demonstrates starkly that collective action driven by robust political consensus is not sufficient for steering complex socio-technical systems in desired directions.
ContributorsSturm, Christine (Author) / Sarewitz, Daniel (Thesis advisor) / Miller, Clark (Committee member) / Anderies, John (Committee member) / Hirt, Paul (Committee member) / Arizona State University (Publisher)
Created2018
Description

We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at

We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using x-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ~38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before-destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods.

ContributorsHunter, Mark S. (Author) / Segelke, Brent (Author) / Messerschmidt, Marc (Author) / Williams, Garth J. (Author) / Zatsepin, Nadia (Author) / Barty, Anton (Author) / Benner, W. Henry (Author) / Carlson, David B. (Author) / Coleman, Matthew (Author) / Graf, Alexander (Author) / Hau-Riege, Stefan P. (Author) / Pardini, Tommaso (Author) / Seibert, M. Marvin (Author) / Evans, James (Author) / Boutet, Sebastien (Author) / Frank, Matthias (Author) / College of Liberal Arts and Sciences (Contributor)
Created2014-08-12
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Description

The context in which many self-governed commons systems operate will likely be significantly altered as globalization processes play out over the next few decades. Such dramatic changes will induce some systems to fail and subsequently to be transformed, rather than merely adapt. Despite this possibility, research on globalization-induced transformations of

The context in which many self-governed commons systems operate will likely be significantly altered as globalization processes play out over the next few decades. Such dramatic changes will induce some systems to fail and subsequently to be transformed, rather than merely adapt. Despite this possibility, research on globalization-induced transformations of social-ecological systems (SESs) is still underdeveloped. We seek to help fill this gap by exploring some patterns of transformation in SESs and the question of what factors help explain the persistence of cooperation in the use of common-pool resources through transformative change. Through the analysis of 89 forest commons in South Korea that experienced such transformations, we found that there are two broad types of transformation, cooperative and noncooperative. We also found that two system-level properties, transaction costs associated group size and network diversity, may affect the direction of transformation. SESs with smaller group sizes and higher network diversity may better organize cooperative transformations when the existing system becomes untenable.

ContributorsYu, David (Author) / Anderies, John (Author) / Lee, Dowon (Author) / Perez, Irene (Author) / Julie Ann Wrigley Global Institute of Sustainability (Contributor)
Created2013-11-30
Description

Photosynthesis, a process catalysed by plants, algae and cyanobacteria converts sunlight to energy thus sustaining all higher life on Earth. Two large membrane protein complexes, photosystem I and II (PSI and PSII), act in series to catalyse the light-driven reactions in photosynthesis. PSII catalyses the light-driven water splitting process, which

Photosynthesis, a process catalysed by plants, algae and cyanobacteria converts sunlight to energy thus sustaining all higher life on Earth. Two large membrane protein complexes, photosystem I and II (PSI and PSII), act in series to catalyse the light-driven reactions in photosynthesis. PSII catalyses the light-driven water splitting process, which maintains the Earth’s oxygenic atmosphere. In this process, the oxygen-evolving complex (OEC) of PSII cycles through five states, S0 to S4, in which four electrons are sequentially extracted from the OEC in four light-driven charge-separation events. Here we describe time resolved experiments on PSII nano/microcrystals from Thermosynechococcus elongatus performed with the recently developed technique of serial femtosecond crystallography. Structures have been determined from PSII in the dark S1 state and after double laser excitation (putative S3 state) at 5 and 5.5 Å resolution, respectively. The results provide evidence that PSII undergoes significant conformational changes at the electron acceptor side and at the Mn4CaO5 core of the OEC. These include an elongation of the metal cluster, accompanied by changes in the protein environment, which could allow for binding of the second substrate water molecule between the more distant protruding Mn (referred to as the ‘dangler’ Mn) and the Mn3CaOx cubane in the S2 to S3 transition, as predicted by spectroscopic and computational studies. This work shows the great potential for time-resolved serial femtosecond crystallography for investigation of catalytic processes in biomolecules.

ContributorsKupitz, Christopher (Author) / Basu, Shibom (Author) / Grotjohann, Ingo (Author) / Fromme, Raimund (Author) / Zatsepin, Nadia (Author) / Rendek, Kimberly (Author) / Hunter, Mark (Author) / Shoeman, Robert L. (Author) / White, Thomas A. (Author) / Wang, Dingjie (Author) / James, Daniel (Author) / Yang, Jay-How (Author) / Cobb, Danielle (Author) / Reeder, Brenda (Author) / Sierra, Raymond G. (Author) / Liu, Haiguang (Author) / Barty, Anton (Author) / Aquila, Andrew L. (Author) / Deponte, Daniel (Author) / Kirian, Richard (Author) / Bari, Sadia (Author) / Bergkamp, Jesse (Author) / Beyerlein, Kenneth R. (Author) / Bogan, Michael J. (Author) / Caleman, Carl (Author) / Chao, Tzu-Chiao (Author) / Conrad, Chelsie (Author) / Davis, Katherine M. (Author) / Department of Chemistry and Biochemistry (Contributor)
Created2014-09-11
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Description

The insights in Governing the Commons have provided foundational ideas for commons research in the past 23 years. However, the cases that Elinor Ostrom analyzed have been exposed to new social, economic, and ecological disturbances. What has happened to these cases since the 1980s? We reevaluated one of Ostrom’s case

The insights in Governing the Commons have provided foundational ideas for commons research in the past 23 years. However, the cases that Elinor Ostrom analyzed have been exposed to new social, economic, and ecological disturbances. What has happened to these cases since the 1980s? We reevaluated one of Ostrom’s case studies, the lobster and groundfishery of Port Lameron, Southwest Nova Scotia (SWNS). Ostrom suggested that the self-governance of this fishery was fragile because the government did not recognize the rights of resource users to organize their own rules. In the Maine lobster fishery, however, the government formalized customary rules and decentralized power to fishing ports. We applied the concepts of feedback, governance mismatches, and the robustness of social-ecological systems to understand the pathway of institutional change in Port Lameron. We revisited the case of Port Lameron using marine harvesters’ accounts collected from participant observation, informal interviews and surveys, and literature on fisheries policy and ecology in SWNS and Maine. We found that the government’s failure to recognize the customary rights of harvesters to organize has weakened feedback between the operational level, where resource users interact with the resource, and the collective-choice level, where agents develop rules to influence the behavior of resource users. This has precipitated governance mismatches, which have led harvesters to believe that the decision-making process is detrimental to their livelihoods. Thus, harvesters rarely participate in decision making and resist regulatory change. In Maine, harvesters can influence decisions through participation, but there is a trade-off. With higher influence in decisions, captains have co-opted the decision-making process. Nevertheless, we suggest that the fisheries of SWNS are more vulnerable to social-ecological change because of weaker feedbacks than in Maine. Finally, we have discussed the potential benefits of polycentricity to both fisheries.

ContributorsBarnett, Allain (Author) / Anderies, John (Author) / College of Liberal Arts and Sciences (Contributor)
Created2013-11-30
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Description

It has been suggested that the extended intensity profiles surrounding Bragg reflections that arise when a series of finite crystals of varying size and shape are illuminated by the intense, coherent illumination of an x-ray free-electron laser may enable the crystal’s unit-cell electron density to be obtained ab initio via

It has been suggested that the extended intensity profiles surrounding Bragg reflections that arise when a series of finite crystals of varying size and shape are illuminated by the intense, coherent illumination of an x-ray free-electron laser may enable the crystal’s unit-cell electron density to be obtained ab initio via well-established iterative phasing algorithms. Such a technique could have a significant impact on the field of biological structure determination since it avoids the need for a priori information from similar known structures, multiple measurements near resonant atomic absorption energies, isomorphic derivative crystals, or atomic-resolution data. Here, we demonstrate this phasing technique on diffraction patterns recorded from artificial two-dimensional microcrystals using the seeded soft x-ray free-electron laser FERMI. We show that the technique is effective when the illuminating wavefront has nonuniform phase and amplitude, and when the diffraction intensities cannot be measured uniformly throughout reciprocal space because of a limited signal-to-noise ratio.

ContributorsKirian, Richard (Author) / Bean, Richard J. (Author) / Beyerlein, Kenneth R. (Author) / Barthelmess, Miriam (Author) / Yoon, Chun Hong (Author) / Wang, Fenglin (Author) / Capotondi, Flavio (Author) / Pedersoli, Emanuele (Author) / Barty, Anton (Author) / Chapman, Henry N. (Author) / College of Liberal Arts and Sciences (Contributor)
Created2015-02-12
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Description

What's a profession without a code of ethics? Being a legitimate profession almost requires drafting a code and, at least nominally, making members follow it. Codes of ethics (henceforth “codes”) exist for a number of reasons, many of which can vary widely from profession to profession - but above all

What's a profession without a code of ethics? Being a legitimate profession almost requires drafting a code and, at least nominally, making members follow it. Codes of ethics (henceforth “codes”) exist for a number of reasons, many of which can vary widely from profession to profession - but above all they are a form of codified self-regulation. While codes can be beneficial, it argues that when we scratch below the surface, there are many problems at their root. In terms of efficacy, codes can serve as a form of ethical window dressing, rather than effective rules for behavior. But even more that, codes can degrade the meaning behind being a good person who acts ethically for the right reasons.

Created2013-11-30