LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks
Membrane proteins make up approximately a third of all proteins and serve as prime drug targets, yet their structural and biophysical characterization presents some unique challenges due to their hydrophobic surfaces and flexible structure. The main goal of this project was to compare, with the use...
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Umeå universitet, Kemiska institutionen
2021
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ndltd-UPSALLA1-oai-DiVA.org-umu-1855472021-07-01T05:24:57ZLMCA1 as prototype system for the study of Ca2+ pumps in nanodisksengMagkakis, KonstantinosUmeå universitet, Kemiska institutionen2021Chemical SciencesKemiMembrane proteins make up approximately a third of all proteins and serve as prime drug targets, yet their structural and biophysical characterization presents some unique challenges due to their hydrophobic surfaces and flexible structure. The main goal of this project was to compare, with the use of activity assays, two methods employed for the experimental handling of membrane proteins. The methods compared were the traditional method of using detergent micelles to keep the membrane proteins in solution, with the novel technique of reconstituting the proteins in nanodisks, using the LMCA1 P-type ATPase as a prototype. The secondary goal was to successfully express, purify and reconstitute in nanodisks, an inactive mutant of the LMCA1 protein to use as a control for the activity measurements. As of the end of this project the inactive LMCA1 mutant was successfully reconstituted into MSP1D1 nanodisks and the initial assay results showed higher activity for the reconstituted protein. This points to the conclusion that nanodisks could provide an environment that is more native for the protein, compared to detergent micelles, which can lead to greater precision in their structural and biophysical characterization. Student thesisinfo:eu-repo/semantics/bachelorThesistexthttp://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-185547application/pdfinfo:eu-repo/semantics/openAccess |
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Others
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Chemical Sciences Kemi Magkakis, Konstantinos LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
description |
Membrane proteins make up approximately a third of all proteins and serve as prime drug targets, yet their structural and biophysical characterization presents some unique challenges due to their hydrophobic surfaces and flexible structure. The main goal of this project was to compare, with the use of activity assays, two methods employed for the experimental handling of membrane proteins. The methods compared were the traditional method of using detergent micelles to keep the membrane proteins in solution, with the novel technique of reconstituting the proteins in nanodisks, using the LMCA1 P-type ATPase as a prototype. The secondary goal was to successfully express, purify and reconstitute in nanodisks, an inactive mutant of the LMCA1 protein to use as a control for the activity measurements. As of the end of this project the inactive LMCA1 mutant was successfully reconstituted into MSP1D1 nanodisks and the initial assay results showed higher activity for the reconstituted protein. This points to the conclusion that nanodisks could provide an environment that is more native for the protein, compared to detergent micelles, which can lead to greater precision in their structural and biophysical characterization. |
author |
Magkakis, Konstantinos |
author_facet |
Magkakis, Konstantinos |
author_sort |
Magkakis, Konstantinos |
title |
LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
title_short |
LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
title_full |
LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
title_fullStr |
LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
title_full_unstemmed |
LMCA1 as prototype system for the study of Ca2+ pumps in nanodisks |
title_sort |
lmca1 as prototype system for the study of ca2+ pumps in nanodisks |
publisher |
Umeå universitet, Kemiska institutionen |
publishDate |
2021 |
url |
http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-185547 |
work_keys_str_mv |
AT magkakiskonstantinos lmca1asprototypesystemforthestudyofca2pumpsinnanodisks |
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1719415100936290304 |