Vývoj technik pulsního značení pro studium dynamiky proteinových komplexů.

Fiala, Jan

(IN ENGLISH) Structural mass spectrometry (MS) is an evolving field of structural biology introducing novel techniques for the characterization of biomolecules. Although MS-based techniques only can provide "low-resolution" information compared to standard high-resolution techniques representing by X-ray crystallography, cryo-electron microscopy or nuclear magnetic resonance, its uniqueness lies in the ability to easily obtain structural information about various biomolecules in their native or native-like environment. By employing various approaches, from protein covalent labelling through chemical cross- linking to ion mobility, structural MS provides insight into the structure and dynamics of proteins and their complexes over a broad timescale. This thesis is dedicated to the development of novel structural MS approaches based on pulse covalent labelling and chemical cross-linking. Employing the developed quench-flow microfluidics apparatus, we performed footprinting experiments on proteins and protein complexes in timescale from a few microseconds to single seconds. Specifically, fast photochemical oxidation of proteins (FPOP) and novel fast fluoro alkylation of proteins (FFAP) techniques were utilized to track structural changes of myoglobin upon release of the prosthetic heme group....

guest :: login Digital Repository
		Search		Submit		Help		About