In-Cell NMR in Mammalian Cells: Part 2 View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2012

AUTHORS

Beata Bekei , Honor May Rose , Michaela Herzig , Philipp Selenko

ABSTRACT

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). More... »

PAGES

55-66

References to SciGraph publications

  • 2010. Transfection of siRNAs in Multiple Myeloma Cell Lines in RNA INTERFERENCE
  • 2008-02. Bacterial pore-forming toxins: The (w)hole story? in CELLULAR AND MOLECULAR LIFE SCIENCES
  • 2005-06. An emergency response team for membrane repair in NATURE REVIEWS MOLECULAR CELL BIOLOGY
  • 1993-09. A guide to the use of pore-forming toxins for controlled permeabilization of cell membranes in MEDICAL MICROBIOLOGY AND IMMUNOLOGY
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/978-1-61779-927-3_5

    DOI

    http://dx.doi.org/10.1007/978-1-61779-927-3_5

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1044097031

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/22760312


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