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Characterizing the Structure and Mechanics of 2D Clathrin Lattices with Atomic Force Microscopy

dc.contributor.advisorSchaap, Iwan Dr.
dc.contributor.authorPlaten, Mitja
dc.date.accessioned2015-10-30T09:27:21Z
dc.date.available2015-10-30T09:27:21Z
dc.date.issued2015-10-30
dc.identifier.urihttp://hdl.handle.net/11858/00-1735-0000-0023-9665-1
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-5341
dc.language.isoengde
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc571.4de
dc.titleCharacterizing the Structure and Mechanics of 2D Clathrin Lattices with Atomic Force Microscopyde
dc.typedoctoralThesisde
dc.contributor.refereeSchaap, Iwan Dr.
dc.date.examination2015-10-22
dc.description.abstractengClathrin is a self-assembling protein involved in intracellular trafficking. Since its discovery in the early 1960s, much has been learned about its structure, assembly, and regulation mechanisms. The three legged monomers, called triskelia, are known to form cages in order to invaginate the cell's plasma membrane and, thus, create coated vesicles which contain incorporated extracellular molecules. So far, clathrin research has mostly been focused on these clathrin cages and clathrin-coated vesicles. However, already in the 1980s researchers observed another form of clathrin assembly, namely flat hexagonal lattices attached to the plasma membrane. Little is known about these structures, and whether these flat lattices are involved in clathrin cage formation has been highly debated. The atomic force microscope (AFM) allows the study of biological samples in liquid and, therefore, near their native state. In this work, we took advantage of this unique capability of the AFM to study the 3D structure of flat clathrin lattices at nanometer resolution. The susceptibility of the lattice to mechanical forces, a common issue when investigating soft biomaterials with AFM, was tested using different scan techniques exerting scanning forces from tens of pN to several nN. After discussing the assembly of the flat clathrin lattice itself, the first part of this thesis shows how the AFM can be used to study biological questions in the clathrin field. First, a highly featured image of a hexagonal lattice pore was reconstructed from multiple low force AFM scans. The obtained image was used to investigate the triskelion orientation inside the lattice, and to determine a change in the pucker angle when compared to triskelia in a cage. Secondly, force maps were acquired to investigate the mechanical influence of the clathrin light chains (CLC) on the stability of the lattice. The findings suggest that the CLC has a role in rigidifying the structure of the triskelia. Beyond the investigation of biological properties and mechanisms of clathrin, this thesis additionally explores the application of the flat lattices in bionanotechnological designs. Besides the very regular spacing of the 30 nm-sized pores, clathrin assemblies provide the possibility of specific functionalization, which turns them into useful matrices for sensors and biosynthetic reactors. Towards this goal, we were able to show that the lattice can be assembled on various surface materials and can be stabilized into durable matrices. AFM experiments showed that the stabilized lattice structure remains intact even after a dehydration and rehydration cycle. Finally, a first demonstration to functionalize the lattice with inorganic particles and biologically active molecules is presented.de
dc.contributor.coRefereeTittmann, Kai Prof. Dr.
dc.subject.engAtomic Force Microscopyde
dc.subject.engAFMde
dc.subject.engClathrinde
dc.subject.engclathrin latticede
dc.subject.engBionanotechnologyde
dc.subject.engnanotechnologyde
dc.subject.engself-assemblyde
dc.subject.engprotein assemblyde
dc.identifier.urnurn:nbn:de:gbv:7-11858/00-1735-0000-0023-9665-1-7
dc.affiliation.instituteGöttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)de
dc.subject.gokfullBiologie (PPN619462639)de
dc.identifier.ppn838132804


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