Activity Assays for 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%3A00483419" target="_blank" >RIV/61388963:_____/17:00483419 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1016/bs.mie.2016.11.002" target="_blank" >http://dx.doi.org/10.1016/bs.mie.2016.11.002</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/bs.mie.2016.11.002" target="_blank" >10.1016/bs.mie.2016.11.002</a>
Alternative languages
Result language
angličtina
Original language name
Activity Assays for Rhomboid Proteases
Original language description
Rhomboids are ubiquitous intramembrane serine proteases that are involved in various signaling pathways. This fascinating class of proteases harbors an active site buried within the lipid milieu. High-resolution structures of the Escherichia coli rhomboid GlpG with various inhibitors revealed the catalytic mechanism for rhomboid-mediated proteolysis, however, a quantitative characterization was lacking. Assessing an enzyme's catalytic parameters is important for understanding the details of its proteolytic reaction and regulatory mechanisms. To assay rhomboid protease activity, many challenges exist such as the lipid environment and lack of known substrates. Here, we summarize various enzymatic assays developed over the last decade to study rhomboid protease activity. We present detailed protocols for gel-shift and FRET-based assays, and calculation of KM and V-max to measure catalytic parameters, using detergent solubilized rhomboids with TatA, the only known substrate for bacterial rhomboids, and the model substrate fluorescently labeled casein.
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
43
Pages from-to
395-437
Number of pages of the book
474
Publisher name
Academic Press
Place of publication
Cambridge
UT code for WoS chapter
000403271000016