Production of Recombinant Rhomboid Proteases
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F17%3A00483405" target="_blank" >RIV/61388963:_____/17:00483405 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1016/bs.mie.2016.10.031" target="_blank" >http://dx.doi.org/10.1016/bs.mie.2016.10.031</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/bs.mie.2016.10.031" target="_blank" >10.1016/bs.mie.2016.10.031</a>
Alternative languages
Result language
angličtina
Original language name
Production of Recombinant Rhomboid Proteases
Original language description
Rhomboid proteases are intramembrane enzymes that hydrolyze peptide bonds of transmembrane proteins in the lipid bilayer. They play a variety of roles in key biological events and are linked to several disease states. Over the last decade a great deal of structural and functional knowledge has been generated on this fascinating class of proteases. Both structural and kinetic analyses require milligram amounts of protein, which may be challenging for membrane proteins such as rhomboids. Here, we present a detailed protocol for optimization of expression and purification of three rhomboid proteases from Escherichia coli (ecGlpG), Haemophilus influenzae (hiGlpG), and Providencia stuartii (AarA). We discuss the optimization of expression conditions, such as concentration of inducing agent, induction time, and temperature, as well as purification protocol with precise details for each step. The provided protocol yields 1-2.5 mg of rhomboid enzyme per liter of bacterial culture and can assist in structural and functional studies of intramembrane proteases.
Czech name
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Czech description
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Classification
Type
C - Chapter in a specialist book
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Book/collection name
Enzymology at the Membrane Interface: Intramembrane Proteases
ISBN
978-0-12-812213-6
Number of pages of the result
24
Pages from-to
255-278
Number of pages of the book
474
Publisher name
Academic Press
Place of publication
Cambridge
UT code for WoS chapter
000403271000011