Chapter title |
In-cell NMR in Mammalian cells: part 2.
|
---|---|
Chapter number | 5 |
Book title |
Intrinsically Disordered Protein Analysis
|
Published in |
Methods in molecular biology, July 2012
|
DOI | 10.1007/978-1-61779-927-3_5 |
Pubmed ID | |
Book ISBNs |
978-1-61779-926-6, 978-1-61779-927-3
|
Authors |
Bekei B, Rose HM, Herzig M, Selenko P, 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). |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Japan | 1 | 5% |
United Kingdom | 1 | 5% |
Germany | 1 | 5% |
Unknown | 19 | 86% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 5 | 23% |
Student > Ph. D. Student | 5 | 23% |
Student > Postgraduate | 3 | 14% |
Other | 2 | 9% |
Professor > Associate Professor | 2 | 9% |
Other | 2 | 9% |
Unknown | 3 | 14% |
Readers by discipline | Count | As % |
---|---|---|
Chemistry | 7 | 32% |
Agricultural and Biological Sciences | 6 | 27% |
Biochemistry, Genetics and Molecular Biology | 5 | 23% |
Engineering | 1 | 5% |
Unknown | 3 | 14% |