Matches in SemOpenAlex for { <https://semopenalex.org/work/W633018934> ?p ?o ?g. }
Showing items 1 to 75 of
75
with 100 items per page.
- W633018934 abstract "This thesis aimed to design novel sensor proteins that can identify and measure various metal ions in vivo and in situ. Metal ions play key role in the metabolism of the cell, and monitoring of calcium has helped interrogate cellular processes such as fertilisation, contraction and apoptosis. Real-time monitoring of more divalent metal ions like zinc and copper is required to gain much needed insight into brain function and associated disorders, such as Alzheimer’s and Parkinson’s disease. Aequorin is a calcium-regulated photoprotein originally isolated from the jellyfish Aequorea victoria. Due to its high sensitivity to calcium and its non-invasive nature, aequorin has been used as a real-time indicator of calcium ions in biological systems for more than forty years. The protein complex consists of the polypeptide chain apoaequorin and a tightly bound chromophore (coelenterazine). Trace amounts of calcium ions trigger conformational changes in the protein, which in turn facilitate the intermolecular oxidation of coelenterazine and concomitant production of CO2 and a flash of blue light. Aequorin’s light emitting reaction can also be triggered by a range of other divalent and trivalent cations, leading however to significantly lower light yields. Based on aequorin’s promiscuity towards other ions, this project tested the hypothesis that aequorin’s preference for certain cations could be manipulated through mutations engineered in one or more of the three calcium-binding loops (EF-hands). In order to test the hypothesis, the following six stages were performed: cloning of the apoaequorin gene for expression in E. coli; development of a high-throughput assay for expression and measurement of bioluminescent activity in microwells; design of a library containing forty eight mutant variants of aequorin; screening of the library against seven metal ions; protein purification of wild-type aequorin and one selected mutant; analysis of activity and kinetics of purified wild type and one chosen mutant against all seven ions. This work produced mutants with shifted selectivity towards new metal ions at the cost of luminescence yield. The impact of mutations is analysed and it is suggested that one of the EF-hands (EF-I) is likely to serve as a gatekeeper to aequorin’s selectivity. It was also shown that at least one mutant utilised zinc ions (that wild type failed to utilise) to achieve low levels of bioluminescent activity." @default.
- W633018934 created "2016-06-24" @default.
- W633018934 creator A5051718560 @default.
- W633018934 date "2014-11-28" @default.
- W633018934 modified "2023-09-26" @default.
- W633018934 title "Redesign of calcium-regulated protein aequorin towards the development of a novel ion bioreporter" @default.
- W633018934 hasPublicationYear "2014" @default.
- W633018934 type Work @default.
- W633018934 sameAs 633018934 @default.
- W633018934 citedByCount "0" @default.
- W633018934 crossrefType "dissertation" @default.
- W633018934 hasAuthorship W633018934A5051718560 @default.
- W633018934 hasConcept C104317684 @default.
- W633018934 hasConcept C122229402 @default.
- W633018934 hasConcept C12554922 @default.
- W633018934 hasConcept C142613039 @default.
- W633018934 hasConcept C17189327 @default.
- W633018934 hasConcept C178790620 @default.
- W633018934 hasConcept C185592680 @default.
- W633018934 hasConcept C192562407 @default.
- W633018934 hasConcept C2777105759 @default.
- W633018934 hasConcept C2985930086 @default.
- W633018934 hasConcept C42086223 @default.
- W633018934 hasConcept C49040817 @default.
- W633018934 hasConcept C5176359 @default.
- W633018934 hasConcept C519063684 @default.
- W633018934 hasConcept C55493867 @default.
- W633018934 hasConcept C74884574 @default.
- W633018934 hasConcept C80933277 @default.
- W633018934 hasConcept C86803240 @default.
- W633018934 hasConceptScore W633018934C104317684 @default.
- W633018934 hasConceptScore W633018934C122229402 @default.
- W633018934 hasConceptScore W633018934C12554922 @default.
- W633018934 hasConceptScore W633018934C142613039 @default.
- W633018934 hasConceptScore W633018934C17189327 @default.
- W633018934 hasConceptScore W633018934C178790620 @default.
- W633018934 hasConceptScore W633018934C185592680 @default.
- W633018934 hasConceptScore W633018934C192562407 @default.
- W633018934 hasConceptScore W633018934C2777105759 @default.
- W633018934 hasConceptScore W633018934C2985930086 @default.
- W633018934 hasConceptScore W633018934C42086223 @default.
- W633018934 hasConceptScore W633018934C49040817 @default.
- W633018934 hasConceptScore W633018934C5176359 @default.
- W633018934 hasConceptScore W633018934C519063684 @default.
- W633018934 hasConceptScore W633018934C55493867 @default.
- W633018934 hasConceptScore W633018934C74884574 @default.
- W633018934 hasConceptScore W633018934C80933277 @default.
- W633018934 hasConceptScore W633018934C86803240 @default.
- W633018934 hasLocation W6330189341 @default.
- W633018934 hasOpenAccess W633018934 @default.
- W633018934 hasPrimaryLocation W6330189341 @default.
- W633018934 hasRelatedWork W1539815641 @default.
- W633018934 hasRelatedWork W1978451197 @default.
- W633018934 hasRelatedWork W1987800116 @default.
- W633018934 hasRelatedWork W2006011889 @default.
- W633018934 hasRelatedWork W20155870 @default.
- W633018934 hasRelatedWork W2019257137 @default.
- W633018934 hasRelatedWork W2057363169 @default.
- W633018934 hasRelatedWork W2068927073 @default.
- W633018934 hasRelatedWork W2098183914 @default.
- W633018934 hasRelatedWork W2111523408 @default.
- W633018934 hasRelatedWork W2127730547 @default.
- W633018934 hasRelatedWork W2156014966 @default.
- W633018934 hasRelatedWork W2181366372 @default.
- W633018934 hasRelatedWork W2418288264 @default.
- W633018934 hasRelatedWork W2560205581 @default.
- W633018934 hasRelatedWork W2737867512 @default.
- W633018934 hasRelatedWork W2805640549 @default.
- W633018934 hasRelatedWork W989798991 @default.
- W633018934 hasRelatedWork W2196699411 @default.
- W633018934 hasRelatedWork W3091789219 @default.
- W633018934 isParatext "false" @default.
- W633018934 isRetracted "false" @default.
- W633018934 magId "633018934" @default.
- W633018934 workType "dissertation" @default.