TY  - JOUR
A1  - Parcej, David
A1  - Guntrum, Renate
A1  - Schmidt, Sabine
A1  - Hinz, Andreas
A1  - Tampé, Robert
T1  - Multicolour fluorescence-detection size-exclusion chromatography for structural genomics of membrane multiprotein complexes
T2  - PLoS One
N2  - Many interesting and important membrane proteins are hetero-oligomeric. However, besides naturally abundant examples, the structures of relatively few such complexes are known. Partly, this is due to difficulties in expression, stoichiometric assembly, and in the evaluation of their stability prior to crystallization trials. Here we describe a new approach, which allows rapid assessment of protein complex quality, assembly and stoichiometry, simplifying the search for conditions conducive to long-term stability and crystallization. Multicolour fluorescence size-exclusion chromatography (MC-FSEC) is used to enable tracking of individual subunits through expression, solubilization and purification steps. We show how the method has been applied to the heterodimeric transporter associated with antigen processing (TAP) and demonstrate how it may be extended in order to analyse membrane multisubunit assemblies.
Y1  - 2013
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/30590
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-305907
SN  - 1932-6203
N1  - Copyright: © 2013 Parcej et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
VL  - 8
IS  - (6):e67112
PB  - PLoS
CY  - Lawrence, Kan.
ER  -