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You searched for subject:(lipidic cubic phase). Showing records 1 – 2 of 2 total matches.

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1. Safari, Cecilia. Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase.

Degree: 2019, University of Gothenburg / Göteborgs Universitet

Life in all living organisms depend on chemical processes performed by proteins, which are molecular machineries encoded in the DNA. Cytochrome c oxidase (CcO) is a membrane protein essential for cellular respiration, a process in which chemical energy enters the electron transport chain in the form of electrons and is transformed to ATP via a proton gradient. In the active site of CcO a redox-reaction takes place, where oxygen that we breathe is reduced to two water molecules at the same time as protons are pumped over the membrane. This thesis focuses on structural investigations performed by serial crystallography (SX) of CcO in lipidic cubic phase (LCP). The advances of bright X-ray sources in the form of X-ray Free Electron Lasers with short pulse durations and 4:th generation synchrotrons have evolved the structural biology field in the sense that high-quality data can be collected at room temperature with minimal X-ray induced radiation damage. These advancements enable the capturing of structural intermediates of proteins in a time-resolved manner. Well-diffracting microcrystals are of essence in serial crystallography experiments, and the study of membrane proteins in lipidic cubic phase has proven to be favorable compared to detergent-based crystallization. This thesis focuses on developing procedures to produce microcrystals in LCP, and its use in studies of ba3-type CcO from Thermus thermophilus as well as reaction centre from Blastrochloris viridis. The room-temperature structure of ba3-type CcO is determined in both the resting oxidized state and the reduced CO-bound state. The resting state structure reveals the active site ligand as a single oxygen species, and in comparisons to previously published structures of CcO we show structural differences between ba3-type CcO and bovine aa3-type CcO upon CO-binding. The work presented in this thesis provides the groundwork for future time-resolved CO-photolysis experiments of ba3-type CcO by the pump-probe approach, which may reveal more about the structural mechanisms that explain proton pumping in CcO.

Subjects/Keywords: Serial crystallography; Cytochrome c oxidase; Microcrystallization in lipidic cubic phase

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Safari, C. (2019). Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase. (Thesis). University of Gothenburg / Göteborgs Universitet. Retrieved from http://hdl.handle.net/2077/62148

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Safari, Cecilia. “Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase.” 2019. Thesis, University of Gothenburg / Göteborgs Universitet. Accessed May 09, 2021. http://hdl.handle.net/2077/62148.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Safari, Cecilia. “Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase.” 2019. Web. 09 May 2021.

Vancouver:

Safari C. Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase. [Internet] [Thesis]. University of Gothenburg / Göteborgs Universitet; 2019. [cited 2021 May 09]. Available from: http://hdl.handle.net/2077/62148.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Safari C. Microcrystallization in lipidic cubic phase and serial crystallography studies of cytochrome c oxidase. [Thesis]. University of Gothenburg / Göteborgs Universitet; 2019. Available from: http://hdl.handle.net/2077/62148

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation


University of Gothenburg / Göteborgs Universitet

2. Andersson, Rebecka. Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography.

Degree: 2020, University of Gothenburg / Göteborgs Universitet

Every living organism contain a cell membrane which is an important cell structure with a vast variety of different functions such as cell signaling, transportation and energy production. One of the most important functions is to produce energy for the cell to thrive. In humans and other organisms, oxygen is used as the final electron acceptor to drive reactions that pump protons across the membrane to create an electrochemical proton gradient. This electrochemical proton gradient is then harvested for the production of ATP, the currency of life. Even though the membrane proteins that are responsible for the electrochemical proton gradient belong to one of the most well-studied membrane protein families, there are still mechanisms to be revealed. Two of these mechanisms are proton pumping across the membrane and the route of oxygen to the active site of cytochrome c oxidase, the final enzyme in the respiratory chain that reduces oxygen to water. By using X-ray serial crystallography these mechanisms can be revealed. Previous research has found that membrane protein crystallization is greatly improved if the environment of the protein mimics the native environment. Reconstituting the membrane proteins in a lipidic cubic phase, a membrane mimicking lipid bilayer, increases membrane protein stability and crystal packing. As a result, large volumes of good quality microcrystals for X-ray serial crystallography can be obtained. Our studies present a method that allows for better visualization of the crystallization process of microcrystals in lipidic cubic phase. The method was then used to produce microcrystals of a ba3-type cytochrome c oxidase which resulted in the first room temperature structure at 2.3 Å resolution. This work was extended by a procedure to bind CO to the active site of the protein crystals, a first step for revealing the mechanisms of proton pumping and oxygen migration within the enzyme. The method was also successfully used for other proteins where new crystallization hits were found and optimized. These include sensory rhodopsin II from halophilic archea and reaction centre from Blastochloris viridis. For reaction centre, the method was also used in combination with crystal seeding to create a new procedure for microcrystallization. The work presented in this thesis provides a foundation for further development of serial crystallography and for producing microcrystals for time resolved studies.

Subjects/Keywords: Structural biology; Lipidic Cubic Phase; Cytochrome c oxidase; Serial Crystallography

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Andersson, R. (2020). Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography. (Thesis). University of Gothenburg / Göteborgs Universitet. Retrieved from http://hdl.handle.net/2077/66804

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Andersson, Rebecka. “Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography.” 2020. Thesis, University of Gothenburg / Göteborgs Universitet. Accessed May 09, 2021. http://hdl.handle.net/2077/66804.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Andersson, Rebecka. “Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography.” 2020. Web. 09 May 2021.

Vancouver:

Andersson R. Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography. [Internet] [Thesis]. University of Gothenburg / Göteborgs Universitet; 2020. [cited 2021 May 09]. Available from: http://hdl.handle.net/2077/66804.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Andersson R. Lipidic Cubic Phase Microcrystallization and its Application in Serial Crystallography. [Thesis]. University of Gothenburg / Göteborgs Universitet; 2020. Available from: http://hdl.handle.net/2077/66804

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

.