Original title: Crystallization, structural and modeling studies of photosynthetic membrane protein and water soluble haloalkane dehalogenase
Authors: PRUDNIKOVA, Tatyana
Document type: Doctoral theses
Year: 2011
Language: eng
Abstract: [eng] [cze]
This PhD thesis is devoted to study of a) water soluble protein haloalkane dehalogenase DbeA (wild type enzyme) and its mutant variant DbeA1 from bacteria Bradyrhizobium elkani USDA94 and also b) the inner membrane protein complex of Photosystem II (PSII) from Pisum sativum L. An active site architecture of the haloalkane dehalogenase enzymes DbeA, DbeA1, stabilization by loop insertion in a mutant form, and structure-function relationships of haloalkane dehalogenases have been studied by using experimental approaches protein crystallization and crystallography. The protein crystals have been grown and X-ray diffraction data on these crystals have been collected and 3D molecular structure has been solved for both dehalogenases DbeA and DbeA1 at 2.2 A? resolution, for DbeA crystals soaked with 1-fluoropentane as a substrate at 1.65 A? resolution, and with 1,3-dichloropropane substrate at 2.15 A? resolution. Crystallization and molecular modelling methods were applied on membrane protein PSII core complex from higher plants in order to find optimal conditions for the 3D crystals formation, to establish the crystallization protocol and to study light-induced processes in PSII reaction center. Tato disertac?ni? pra?ce se zaby?va? studiem a) ve vode? rozpustne?ho proteinu haloalkan dehalogenasy DbeA (enzym divoke?ho typu) a jeho mutantni? variantou DbeA1 z bakterii? Bradyrhizobium elkani USDA94 a b) membra?nove?ho komplexu fotosyste?mu II (PSII) z hrachu sete?ho Pisum sativum L. U enzymu? DbeA a DbeA1 z rodiny haloalkan dehalogenas byla studova?na architektura jejich aktivni?ho mi?sta, stabilizace proteinu inzerci? peptidove? smyc?ky u mutantni? formy a strukturne?-funkc?ni? vztahy s pomoci? experimenta?lni?ch strukturni?ch metod proteinove? krystalizace a krystalografie. V obou pr?i?padech enzymu? DbeA a DbeA1 byly vype?stova?ny proteinove? krystaly a na nich name?r?ena? data z rentgenove? difrakce poskytla 3D molekula?rni? struktury s rozlis?eni?m 2.20 A?. Krystaly dehalogenasy DbeA saturovane? substra?tem 1-fluoropentanem poskytly 3D strukturu s rozlis?eni?m 1.65 A? a v pr?i?pade? substra?tu 1,3-dichlorpropanu bylo dosaz?eno rozlis?eni? 2.15 A?. Prostr?ednictvi?m metod molekula?rni?ho modelova?ni? a proteinove? krystalografie byl take? studova?n membra?novy? protein PSII core komplex z vys?s?i?ch rostlin. V ra?mci krystalizac?ni? studie byly hleda?ny optima?lni? krystalizac?ni? podmi?nky pro vznik 3D krystalu? PSII a byl vytvor?en krystalizac?ni? protokol. Modelova?ni? na semiempiricke?m 3D modelu PSII umoz?nilo studovat sve?tlem indukovane? procesy ve fotosynteticke?m reakc?ni?m centru.
Citation: PRUDNIKOVA, Tatyana. Crystallization, structural and modeling studies of photosynthetic membrane protein and water soluble haloalkane dehalogenase. N. Hrady, 2011. disertační práce (Ph.D.). JIHOČESKÁ UNIVERZITA V ČESKÝCH BUDĚJOVICÍCH. ***Ústav fyzikální biologie

Institution: University of South Bohemia in České Budějovice (web)
Document availability information: Fulltext is available in the Digital Repository of University of South Bohemia.
Original record: http://www.jcu.cz/vskp/12629

Permalink: http://www.nusl.cz/ntk/nusl-71479


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Universities and colleges > Public universities > University of South Bohemia in České Budějovice
Academic theses (ETDs) > Doctoral theses