Matching Items (144)
Description
Diacylglycerol kinase catalyses the ATP-dependent conversion of diacylglycerol to phosphatidic acid in the plasma membrane of Escherichia coli. The small size of this integral membrane trimer, which has 121 residues per subunit, means that available protein must be used economically to craft three catalytic and substrate-binding sites centred about the membrane/cytosol interface. How nature has accomplished this extraordinary feat is revealed here in a crystal structure of the kinase captured as a ternary complex with bound lipid substrate and an ATP analogue. Residues, identified as essential for activity by mutagenesis, decorate the active site and are rationalized by the ternary structure. The γ-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane. A catalytic mechanism for this unique enzyme is proposed. The active site architecture shows clear evidence of having arisen by convergent evolution.
ContributorsLi, Dianfan (Author) / Stansfeld, Phillip J. (Author) / Sansom, Mark S. P. (Author) / Keogh, Aaron (Author) / Vogeley, Lutz (Author) / Howe, Nicole (Author) / Lyons, Joseph A. (Author) / Aragao, David (Author) / Fromme, Petra (Author) / Fromme, Raimund (Author) / Basu, Shibom (Author) / Grotjohann, Ingo (Author) / Kupitz, Christopher (Author) / Rendek, Kimberley (Author) / Weierstall, Uwe (Author) / Zatsepin, Nadia (Author) / Cherezov, Vadim (Author) / Liu, Wei (Author) / Bandaru, Sateesh (Author) / English, Niall J. (Author) / Gati, Cornelius (Author) / Barty, Anton (Author) / Yefanov, Oleksandr (Author) / Chapman, Henry N. (Author) / Diederichs, Kay (Author) / Messerschmidt, Marc (Author) / Boutet, Sebastien (Author) / Williams, Garth J. (Author) / Seibert, M. Marvin (Author) / Caffrey, Martin (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2015-12-17
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 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
Description
Antibodies are essential for structural determinations and functional studies of membrane proteins, but antibody generation is limited by the availability of properly-folded and purified antigen. We describe the first application of genetic immunization to a structurally diverse set of membrane proteins to show that immunization of mice with DNA alone produced antibodies against 71% (n = 17) of the bacterial and viral targets. Antibody production correlated with prior reports of target immunogenicity in host organisms, underscoring the efficiency of this DNA-gold micronanoplex approach. To generate each antigen for antibody characterization, we also developed a simple in vitro membrane protein expression and capture method. Antibody specificity was demonstrated upon identifying, for the first time, membrane-directed heterologous expression of the native sequences of the FopA and FTT1525 virulence determinants from the select agent Francisella tularensis SCHU S4. These approaches will accelerate future structural and functional investigations of therapeutically-relevant membrane proteins.
ContributorsHansen, Debra (Author) / Robida, Mark (Author) / Craciunescu, Felicia (Author) / Loskutov, Andrey (Author) / Dorner, Katerina (Author) / Rodenberry, John-Charles (Author) / Wang, Xiao (Author) / Olson, Tien (Author) / Patel, Hetal (Author) / Fromme, Petra (Author) / Sykes, Kathryn (Author) / Biodesign Institute (Contributor) / Innovations in Medicine (Contributor) / Applied Structural Discovery (Contributor) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor)
Created2016-02-24
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. 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
Description
Fashion is an inherently political and reflective medium for the daily ramblings and revolutions of a society. Much of the time the influence is subtle. Silhouettes and fabrics reflect different stances on conservatism, on sex, on the degrees to which we fetishize luxury, and on infinite other attitudes of an era. Other times the influence is extremely direct, with text printed on the clothing that explicitly articulates a current societal dynamic. I began exploring fashion in 2016, as the country had reached an unprecedented and linguistically weaponized divide.
While taking a fashion technology course under the instruction of Galina Mihaleva, I developed a tracksuit incorporating concealed LED displays that are capable of scrolling customizable text on the sides of the garment. I expanded on this futuristic execution of politically charged clothes by utilizing a more realistic application of the LED technology in the Bouis Vuitton project. This project is a collection of six white vinyl bags with semi-flexible LED displays projecting revolutionary slogans through the vinyl textile.
The bags act as an appropriate housing for technology that is intended for significantly longer use, as bags have a longer lifespan in wardrobes than clothes and return to trend more frequently. The production investment in the technology is more equitable to the investment in the production of a bag and facilitates the wearer’s broadcasting of concise messages. The result is a collection of functional, utilitarian pieces with a clean, futuristic look and a mixed modern and vintage silhouette scrolling pro-revolutionary messages.
Broadcasting the knock-off name ‘BOUIS VUITTON’, I’ve inserted only my first initial into the reputable luxury company and paired it with slogans: ‘EAT THE RICH’ and ‘HEADS WILL ROLL’. The collection articulates a sense of nihilism felt by the youngest generations growing up on the outside of a very exclusive economic and political sphere. Three upcycled vintage luggage pieces evoke associations with the white American upper-class society of the 1960s. The luggage pieces were retrofitted in white vinyl and white-enameled metal fixtures. Three additional soft bags made of the same material reflect a utilitarian style of functional bags on trend with Spring/Summer 2019 streetwear. For the runway presentation of the bags, the models are dressed in navy-colored Dickies boiler suits, white retro-style Fila sneakers, and white ascots reminiscent of the historical male ruffled cravat. The contradictions of iconic silhouettes from both upper and lower-class American fashion history further the juxtaposition of anti-capitalist slogans posted on luxury goods.
Bouis Vuitton: Bags for the Revolution is intended to embody an unapologetic disregard for established wealth and political power in the most public of venues: the sidewalk, the mall, the high and the low-income neighborhoods – wherever people are wearing clothes. Fashion is the modern protest that requires no permit, and the new poster is a luxury bag.
While taking a fashion technology course under the instruction of Galina Mihaleva, I developed a tracksuit incorporating concealed LED displays that are capable of scrolling customizable text on the sides of the garment. I expanded on this futuristic execution of politically charged clothes by utilizing a more realistic application of the LED technology in the Bouis Vuitton project. This project is a collection of six white vinyl bags with semi-flexible LED displays projecting revolutionary slogans through the vinyl textile.
The bags act as an appropriate housing for technology that is intended for significantly longer use, as bags have a longer lifespan in wardrobes than clothes and return to trend more frequently. The production investment in the technology is more equitable to the investment in the production of a bag and facilitates the wearer’s broadcasting of concise messages. The result is a collection of functional, utilitarian pieces with a clean, futuristic look and a mixed modern and vintage silhouette scrolling pro-revolutionary messages.
Broadcasting the knock-off name ‘BOUIS VUITTON’, I’ve inserted only my first initial into the reputable luxury company and paired it with slogans: ‘EAT THE RICH’ and ‘HEADS WILL ROLL’. The collection articulates a sense of nihilism felt by the youngest generations growing up on the outside of a very exclusive economic and political sphere. Three upcycled vintage luggage pieces evoke associations with the white American upper-class society of the 1960s. The luggage pieces were retrofitted in white vinyl and white-enameled metal fixtures. Three additional soft bags made of the same material reflect a utilitarian style of functional bags on trend with Spring/Summer 2019 streetwear. For the runway presentation of the bags, the models are dressed in navy-colored Dickies boiler suits, white retro-style Fila sneakers, and white ascots reminiscent of the historical male ruffled cravat. The contradictions of iconic silhouettes from both upper and lower-class American fashion history further the juxtaposition of anti-capitalist slogans posted on luxury goods.
Bouis Vuitton: Bags for the Revolution is intended to embody an unapologetic disregard for established wealth and political power in the most public of venues: the sidewalk, the mall, the high and the low-income neighborhoods – wherever people are wearing clothes. Fashion is the modern protest that requires no permit, and the new poster is a luxury bag.
ContributorsViton, Benjamin Douglas (Author) / Sewell, Dennita (Thesis director) / Mihaleva, Galina (Committee member) / School of Art (Contributor) / College of Integrative Sciences and Arts (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Developments in structural biology has led to advancements in drug design and vaccine development. By better understanding the macromolecular structure, rational choices can be made to improve factors in such as binding affinity, while reducing promiscuity and off-target interactions, improving the medicines of tomorrow. The majority of diseases have a macromolecular basis where rational drug development can make a large impact. Two challenging protein targets of different medical relevance have been investigated at different stages of determining their structures with the ultimate goal of advancing in drug development. The first protein target is the CapBCA membrane protein complex, a virulence factor from the bacterium Francisella tularensis and the causative agent of tularemia and classified as a potential bioterrorism weapon by the United States. Purification of the individual protein targets from the CapBCA complex is a key and challenging step that has been, so far, a limiting factor towards the structure determination of the whole complex. Here, the purification protocols for the CapB and CapC subunits have been establish, which will allow us to progress towards biophysical and structural studies. The second protein target investigated in this thesis is the catalytically active Taspase1. Taspase1 functions as a non-oncogene addiction protease that coordinates cancer cell proliferation and apoptosis and has been found to be overexpressed in many primary human cancers. Here the structure is presented to 3.04A with the goal of rational drug design of Taspase1 inhibitors. Development of Taspase1 inhibitors has no completion in the drug discovery arena and would function as a new anti-cancer therapeutic. Solving the structures of medically relevant proteins such as these is critical towards rapidly developing treatments and prevention of old and new diseases.
ContributorsJernigan, Rebecca J. (Author) / Fromme, Petra (Thesis director) / Hansen, Debra T. (Committee member) / Martin-Garcia, Jose M. (Committee member) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
Structure is a critical component in drug development. This project supports antibody- facilitated structure determination for the following eleven membrane proteins: the human histamine and dopamine G protein-coupled receptors (HRH4 and DRD2) involved in a wide variety of pathologies such as allergies, inflammation, asthma, pain along with Parkinson's and schizophrenia respectively, the human cystic fibrosis transmembrane conductance regulator (CFTR), the human NaV1.8 voltage-gated sodium ion channel, the human TPC2 two-pore channel, the SARS virus proteins 3a, E and M, the MERS virus protein E and M, and the malarial chloroquine resistance transporter (PfCRT). Serum antibodies against these proteins were generated by genetic immunization, and both in vitro and in vivo expressed membrane proteins were created to characterize the serum antibodies. Plasmid clones were generated for genetic immunization, in vitro protein expression, and in vivo expression (HEK293T transfection). Serum antibodies were generated by genetic immunization of mice by gene gun. Genetic immunization promotes an immune response that allows for the generation of antibodies in the absence of purified protein. In vitro expression was accomplished through the novel technique: in vitro translation with hydrophobic magnetic beads (IVT-HMB). Transfections were performed using the HEK293T cell line to express the protein in vivo. The generated protein was then used in gel electrophoresis and silver stain and/or Western blot analyses to identify and visualize the proteins. These expressed proteins will allow for forthcoming characterization of the generated antibodies. The resulting antibodies will in turn enable structure determination of these important membrane proteins by co-crystallization.
ContributorsDrotar, Beniamin (Author) / Fromme, Petra (Thesis director) / Hansen, Debra T. (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
Computer Science and Dance are choice driven disciplines. The output of their processes are compositions of experience. Dancers are not computers and computers are not people but there are comparable traces of humanity in the way each interpret and interact with their respective inputs, outputs, and environments. These overlaps are perhaps not obvious, but in an increasingly specialized world it is important to discuss them. Dynamic Programming and improvisational movement exist within exclusive corners of their respective fields and are characterized by their inherent adaption to change. Inspired by the work of Ivar Hagendoorn, John Cage and other interdisciplinary artists, complexMovement is motivated by the need to create space for intersections between these two powerful groups and find overlaps in the questions they ask to achieve their goals. Dance and Computer Science are just one example of hidden partnerships between their respective fields. Their respective sides allow for ample side by side comparisons but for the purpose of this work, we will focus upon two smaller sectors of their studies: improvisational movement and the design of Dynamic Programming algorithms.
ContributorsOhlsen, Lai Yi Ni (Author) / Britt, Melissa (Thesis director) / Crissman, Angel (Committee member) / Standley, Eileen (Committee member) / Computer Science and Engineering Program (Contributor) / School of Art (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
The topic of my creative project centers on the question of "How can the audience's choices influence dancers' improvisation?" This dance work seeks to redefine the relationship between audience and performers through integration of audience, technology, and movement in real-time. This topic was derived from the fields of Computer Science and Dance. To answer my main question, I need to explore how I can interconnect the theory of Computer Science/fundamentals of a web application and the elements of dance improvisation. This topic interests me because it focuses on combining two studies that do not seem related. However, I find that when I am coding a web application, I can insert blocks of code. This relates to dance improvisation where I have a movement vocabulary, and I can insert different moves based on the context. The idea of gathering data from an audience in real time also interests me. I find that data is most useful when a story can be deduced from that data. To figure out how I can use dance to create and tell a story about the data that is collected, I find that to be intriguing as well. The main goals of my Creative Project are to learn the skills needed to develop a web application using the knowledge and theory that I am acquiring through Computer Science as well as learning about the skills needed to produce a performance piece. My object for the overall project is to create an audience-interactive experience that presents choices for dancers and creates a connection between two completely different studies: Computer Science and Dance. My project will consist of having the audience enter their answers to preset questions via an online voting application. The stage background screen will be utilized to show the question results in percentages in the form of a chart. The dancers will then serve as a live interpretation of these results. This Creative Project will serve as a gateway between the work that has been cultivated in my studies and the real world. The methods involve exploring movement qualities in improvisation, communicating with my cast about what worked best for the transitions between each section of the piece, and testing for the web applications. I learned the importance of having structure within improvisational movement for the purpose of choreography. The significance of structure is that it provides direction, clarity, and a sense of unification for the dancers. I also learned the basics of the programming language, Python, in order to develop the two real-time web applications. The significance of learning Python is that I will be able to add this to my skillset of programming languages as well as build upon my knowledge of Computer Science and develop more real-world applications in the future.
ContributorsNgai, Courtney Taylor (Author) / Britt, Melissa (Thesis director) / Standley, Eileen (Committee member) / Computer Science and Engineering Program (Contributor) / School of Film, Dance and Theatre (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
The purpose of this project was to foster non-inhibited movement exploration such as the movement of untrained dancers in a setting of a music festival (specifically in this case the Grateful Dead community), into the more ritual performance-based ideals and perspectives that tend to occur in a dance studio setting. The external visual perceptions of what an ideal dancer ‘should look like’ lends itself to unrealistic expectations and unattainable goals as an artist. Body image and the lack of individualized self-expression is a problem in studio settings and the goal of this research was to use the perspective of untrained and trained dancers to dig deep into movement qualities that are not contrived or performed from ritual or preconceived notions of movement that tend to occur in trained dancers. Through exploratory improvisational somatic experiences helping the dancer access a more embodied and authentic self, the choreography was shaped through delving into the dancers lived experiences. This study culminated in a performance dance project that was filmed at Arcosanti, an experimental Artist community in Northern Arizona that integrates the design of architecture with a respect to the ecology. The goal is to limit the environmental footprint that is left by the community.
ContributorsFox, Tiffany Suzanne (Author) / Dyer, Becky (Thesis advisor) / Kaplan, Rob (Committee member) / Standley, Eileen (Committee member) / Arizona State University (Publisher)
Created2021