PMID- 18581036
OWN - NLM
STAT- MEDLINE
DCOM- 20081126
LR  - 20220503
IS  - 0022-2631 (Print)
IS  - 0022-2631 (Linking)
VI  - 223
IP  - 1
DP  - 2008 May
TI  - Biophysical and ion channel functional characterization of the Torpedo 
      californica nicotinic acetylcholine receptor in varying detergent-lipid 
      environments.
PG  - 13-26
LID - 10.1007/s00232-008-9107-7 [doi]
AB  - The nicotinic acetylcholine receptor (nAChR) of Torpedo electric rays has been 
      extensively characterized over the last three decades. However, high-resolution 
      structural studies have been hampered by the lack of mechanistic molecular models 
      that describe how detergents influence membrane protein stability and function. 
      Furthermore, elucidation of the dynamic detergent-lipid-protein interactions of 
      solubilized membrane proteins is a largely unexplored research field. This study 
      examines the effects of nine detergents on: (1) nAChR-lipid composition (gas 
      chromatography with flame ionization; GC-FID and/or mass selective detectors; 
      GC-MSD), (2) stability and aggregation state (analytical size exclusion 
      chromatography; A-SEC and electron microscopy; EM) and (3) ion channel function 
      (planar lipid bilayers). Detergent solubilization of nAChR-enriched membranes did 
      not result in significant native lipid depletion or destabilization. Upon 
      purification, native lipid depletion occurred in all detergents, with 
      lipid-analogue detergents CHAPS 
      (3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate, FC-12 
      (n-dodecylphosphocholine) and sodium cholate 
      (3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic acid) maintaining stability 
      and supporting ion channel function, and non-lipid-analogue detergents Cymal-6 
      (6-cyclohexyl-1-hexyl-beta-D-maltoside), DDM (n-dodecyl-beta-D-maltopyranoside), 
      LDAO (lauryldimethylamine-N-oxide) and OG (n-octyl-beta-d-glucopyranoside) 
      decreasing stability and significantly reducing or completely suppressing ion 
      channel function. Anapoe-C(12)E(9 )(polyoxyethylene-[9]-dodecyl ether) and 
      BigCHAP (N,N'-bis-[3-d-gluconamidopropyl] cholamide) retained residual amounts of 
      native lipid, maintaining moderate stability and ion channel function compared to 
      lipid-analogue detergents. Therefore, the nAChR can be stable and functional in 
      lipid-analogue detergents or in detergents that retain moderate amounts of 
      residual native lipids, but not in non-lipid-analogue detergents.
FAU - Asmar-Rovira, Guillermo A
AU  - Asmar-Rovira GA
AD  - Departments of Cell Biology and Molecular Biology, The Scripps Research 
      Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
FAU - Asseo-Garcia, Aloysha M
AU  - Asseo-Garcia AM
FAU - Quesada, Orestes
AU  - Quesada O
FAU - Hanson, Michael A
AU  - Hanson MA
FAU - Cheng, Anchi
AU  - Cheng A
FAU - Nogueras, Carlos
AU  - Nogueras C
FAU - Lasalde-Dominicci, Jose A
AU  - Lasalde-Dominicci JA
FAU - Stevens, Raymond C
AU  - Stevens RC
LA  - eng
GR  - S06 GM 008102/GM/NIGMS NIH HHS/United States
GR  - P41 RR017573-04/RR/NCRR NIH HHS/United States
GR  - P41 RR017573/RR/NCRR NIH HHS/United States
GR  - U54 NS043011/NS/NINDS NIH HHS/United States
GR  - P50 GM073197/GM/NIGMS NIH HHS/United States
GR  - R25 GM061151/GM/NIGMS NIH HHS/United States
GR  - U54 NS043011-06/NS/NINDS NIH HHS/United States
GR  - 2R01 GM 056371-11/GM/NIGMS NIH HHS/United States
GR  - RR 17573/RR/NCRR NIH HHS/United States
GR  - P20 RR016470-02/RR/NCRR NIH HHS/United States
GR  - P20 GM103642/GM/NIGMS NIH HHS/United States
GR  - P20 RR 016470/RR/NCRR NIH HHS/United States
GR  - S06 GM008102/GM/NIGMS NIH HHS/United States
GR  - R01 GM056371-07/GM/NIGMS NIH HHS/United States
GR  - R25 GM 061151/GM/NIGMS NIH HHS/United States
GR  - P50 GM073197-04/GM/NIGMS NIH HHS/United States
GR  - U54 NS 043011/NS/NINDS NIH HHS/United States
GR  - S06 GM008102-340073/GM/NIGMS NIH HHS/United States
GR  - P50 GM 073197/GM/NIGMS NIH HHS/United States
GR  - R01 GM056371/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080625
PL  - United States
TA  - J Membr Biol
JT  - The Journal of membrane biology
JID - 0211301
RN  - 0 (Detergents)
RN  - 0 (Ion Channels)
RN  - 0 (Lipids)
RN  - 0 (Receptors, Nicotinic)
SB  - IM
MH  - Animals
MH  - Biophysical Phenomena
MH  - Detergents/*chemistry
MH  - Dimerization
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Ion Channels/chemistry/physiology/ultrastructure
MH  - Lipids/*chemistry
MH  - Microscopy, Electron
MH  - Models, Chemical
MH  - Molecular Structure
MH  - Receptors, Nicotinic/*chemistry/physiology/ultrastructure
MH  - Torpedo/*metabolism
PMC - PMC2668541
MID - NIHMS97217
EDAT- 2008/06/27 09:00
MHDA- 2008/12/17 09:00
PMCR- 2009/05/01
CRDT- 2008/06/27 09:00
PHST- 2008/02/12 00:00 [received]
PHST- 2008/04/11 00:00 [accepted]
PHST- 2008/06/27 09:00 [pubmed]
PHST- 2008/12/17 09:00 [medline]
PHST- 2008/06/27 09:00 [entrez]
PHST- 2009/05/01 00:00 [pmc-release]
AID - 10.1007/s00232-008-9107-7 [doi]
PST - ppublish
SO  - J Membr Biol. 2008 May;223(1):13-26. doi: 10.1007/s00232-008-9107-7. Epub 2008 
      Jun 25.