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Functional interfaces for the immobilization of membrane proteins : concept, characterization and interactions

Abstract : This thesis is dedicated towards the development of supramolecular assemblies, which are capable of mimicking the amphiphilic nature of the cytoplasmic cell membranes. To this effect, Supported Lipid Bilayers (SLB) was designed to incorporate FhuA (an E.coli outer membrane protein). The interaction of FhuA present in the SLB, with pb5 (the bacteriophage T5 protein), was then studied using QCM-D. Further, Tethered Lipid Bilayer Membranes (tBLM) were constructed on Self-Assembled Monolayers (SAMs) of a novel synthetic anchoring thiol. In this study, the tBLM formation was elaborately investigated using a host of techniques such as QCM-D, AFM and EIS, to infer upon the role of the anchoring thiol in the tBLM formation process. Further, a biotinylated Amphipol (B-PCApol) was employed to immobilize membrane proteins such as FhuA and the human αvβ3 integrin on streptavidin containing surfaces. Using their respective assemblies, the dissociation constant of the FhuA-pb5 complex was determined, whereas the interactions of integrin with its ligand vitronectin were studied by SPR. The last part of this thesis, deals with the study of biomolecular recognition events between a lectin (ConA) and multivalent sugars presented on a RAFT scaffold.
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Hajra Basit. Functional interfaces for the immobilization of membrane proteins : concept, characterization and interactions. Biomolecules [q-bio.BM]. Université de Grenoble, 2011. English. ⟨NNT : 2011GRENV017⟩. ⟨tel-00685163v2⟩

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