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Improving electron microscopic hardware to push the resolution limits in single particle cryo-EM

dc.contributor.advisorStark, Holger Prof. Dr.
dc.contributor.authorYip, Ka Man
dc.titleImproving electron microscopic hardware to push the resolution limits in single particle cryo-EMde
dc.contributor.refereeTittmann, Kai Prof. Dr.
dc.description.abstractengElectron cryomicroscopy (cryo-EM) is a powerful tool in structural biology for determining 3D structures of macromolecular complexes. In the previous decade, many technological advancements have been introduced to improve the methodology of single particle cryo-EM, and hence the resolution of its protein reconstructions. These advancements include developments in both software algorithms and microscope hardware. Despite all these developments, the resolution of single particle cryo-EM cannot surpass 1.5 Å, a barrier blocking direct visualization of atomic details of the protein structure. Here, we show that by improving the optical performance of the electron microscope with new hardware, such resolution barrier can be removed, and atomic resolution of 1.25 Å can be reached with an engineered protein apoferritin. The atomic resolution protein structure helped set benchmarks for validating data quality in high resolution cryo-EM. We found that the new microscope also benefited to other proteins with lower contrast and higher flexibility, such as proteasome, chaperonin and fatty acid synthetase. For these proteins, sample properties and behavior were the major limitations. We anticipate that this would be alleviated by introducing other new microscope hardware, so that in the future, time and effort of structural biologists can concentrate on biochemical preparation of the sample, and single particle cryo-EM can routinely yield high quality protein structures at atomic
dc.contributor.coRefereeHabeck, Michael Dr.
dc.contributor.thirdRefereeEnderlein, Jörg Prof. Dr.
dc.contributor.thirdRefereeDe Groot, Bert Prof. Dr.
dc.contributor.thirdRefereeFaesen, Alexis Caspar Dr.
dc.subject.engsingle particle analysisde
dc.subject.engtransmission electron microscopyde
dc.subject.engaberration correctorde
dc.subject.engenergy filterde
dc.affiliation.instituteBiologische Fakultät für Biologie und Psychologiede
dc.subject.gokfullBiologie (PPN619462639)de
dc.notes.confirmationsentConfirmation sent 2022-07-19T14:45:01de

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