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In-cell NMR in Mammalian cells: part 2.
MedLine Citation:
PMID:  22760312     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Delivery of isotope-labeled IDPs into mammalian cells for the purpose of generating suitable in-cell NMR samples can also be facilitated by action of pore-forming bacterial toxins. In the course of this procedure, mammalian cell membranes are permeated for short periods of time in order to enable the influx of exogenous proteins via a concentration gradient between the outside and the inside of the targeted "host" cells. In contrast to CPP-mediated IDP uptake, toxins offer the advantage that cellular protein transduction does not rely on active biological processes like endocytosis, but on simple passive diffusion. Therefore, proteins that are to be delivered into mammalian cells are not required to contain additional "targeting" sequences, and can be employed in their native contexts. The protocol outlined here employs isotope-labeled human α-synuclein, adherent human HeLa cells, and the Streptococcus pyogenes endotoxin Streptolysin O (SLO).
Authors:
Beata Bekei; Honor May Rose; Michaela Herzig; Philipp Selenko
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  895     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2012  
Date Detail:
Created Date:  2012-07-04     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  55-66     Citation Subset:  IM    
Affiliation:
Department of NMR-assisted Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
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