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dc.contributor.author
Michel, Erich
dc.contributor.author
Allain, Frédéric H.-T.
dc.contributor.editor
Kelman, Zvi
dc.date.accessioned
2022-05-05T06:46:37Z
dc.date.available
2017-06-11T21:13:36Z
dc.date.available
2022-05-05T06:46:37Z
dc.date.issued
2015
dc.identifier.isbn
978-0-12-803048-6
en_US
dc.identifier.issn
0076-6879
dc.identifier.other
10.1016/bs.mie.2015.05.028
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/107011
dc.description.abstract
The steady technical advances of nuclear magnetic resonance (NMR) over the past decades enabled a significant increase in the molecular size of protein particles that can be subjected to a structural and functional characterization in solution. The larger molecular weight of such proteins is accompanied with an increase in NMR signals that complicate spectral interpretation due to signal overlap. The application of segmental isotope labeling to selected domains in multi-domain proteins can significantly facilitate spectral interpretation by reducing the number of observable signals. However, severe signal overlap may persist within individual domains that show low signal dispersion. To further reduce the number of signals and spectral complexity in such systems, we developed a procedure for selective amino acid-type labeling in individual domains of multi-domain proteins. This strategy combines efficient amino acid-type labeling amenable by cell-free protein expression with near-seamless domain ligation achievable by expressed protein ligation. By application of simple dual labeling schemes, this approach further allows residue-specific isotope labeling to position NMR-observable probes at desired sites within segments of multi-domain proteins. This chapter describes a detailed protocol for selective amino acid-type segmental labeling of multi-domain proteins and illustrates its application to a multi-domain RNA-binding protein. The applied ligation approach is further suitable for efficient ligation of unlabeled and/or uniformly labeled domains produced solely by recombinant in vivo expression.
en_US
dc.language.iso
en
en_US
dc.publisher
Academic Press
en_US
dc.subject
Segmental isotope labeling
en_US
dc.subject
Protein ligation
en_US
dc.subject
Amino acid-specific isotope labeling
en_US
dc.subject
Cell-free protein expression
en_US
dc.subject
NMR spectroscopy
en_US
dc.subject
Expressed protein ligation
en_US
dc.title
Selective Amino Acid Segmental Labeling of Multi-Domain Proteins
en_US
dc.type
Book Chapter
dc.date.published
2015-06-18
ethz.book.title
Isotope Labeling of Biomolecules - Labeling Methods
en_US
ethz.journal.title
Methods in Enzymology
ethz.journal.volume
565
en_US
ethz.pages.start
389
en_US
ethz.pages.end
422
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
New York, NY
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
ethz.date.deposited
2017-06-11T21:13:49Z
ethz.source
ECIT
ethz.identifier.importid
imp593653b2e756883513
ethz.ecitpid
pub:167437
ethz.eth
yes
en_US
ethz.availability
Metadata only
en_US
ethz.rosetta.installDate
2017-07-12T20:57:34Z
ethz.rosetta.lastUpdated
2022-03-28T14:17:49Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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