Table 5.6 Some examples of transfer of membrane proteins from amphipols to other amphipols or other
environments.
Amphipol
Final
environment Comments
References
A8-35
Another
APol
An examination of the rate of exchange
of MP-adsorbed APols when exposed to
an excess of a competing APol. In the
case of A8-35, the rate of exchange is
strongly dependent on the ionic strength
Zoonens et al. (2007)
A8-35, A8-75,
OAPA-20,
PMAL-B
Detergent or
mixed
detergent/
lipid micelles
APols can be efficiently and very rapidly
displaced from the TM surface of MPs
by an excess of detergent (cf. Fig. 5.40)
Tribet et al. (1997, 2009), Nagy et al.
(2001), and Zoonens et al. (2007)
OAPA-20,
PMAL-B,
A8-35,
NAPols
Lipid
vesicles
Transfer was initiated by directly
exposing protein/APol complexes to
preformed lipid vesicles. The first study
deals with an α-helical MP,
diacylglycerol kinase (DAGK), the
second one with the pore-forming
domain of diphtheria toxin
Nagy et al. (2001) and Kyrychenko
et al. (2012)
A8-35
Black lipid
film
The functionality of OmpA and FomA
following folding in A8-35 was
examined by applying the complexes to
black lipid films. It was noted that
recovery of native-like pore conductance
requires that the APol be applied to both
the cis and trans sides of the film
(cf. Fig. 5.35)
Pocanschi et al. (2006b)
A8-35
Live cells
A rhodamine-labeled peptide mimicking
the transmembrane anchor of neuropilin1 was trapped with NBD-labeled A8-35
and the distribution of the two species
following delivery to COS-7 cells
followed by confocal fluorescence
microscopy. See Chap. 15, Fig. 15.23
Popot et al. (2011)
A8-35
Lipidic
mesophase
Direct transfer of BR from A8-35 to a
lipidic mesophase led to the formation of
highly ordered BR crystals containing
no APol. See Chap. 11, § 11.3.2
Polovinkin et al. (2014b)
SMA
Lipidic
mesophase
Same as above, using SMALP-trapped
bacteriorhodopsin from Haloquadratum
walsbyi
Broecker et al. (2017)
A8-35
Nanodiscs
Two GPCRs were expressed in E. coli as
inclusion bodies, solubilized in SDS,
folded in A8-35, and transferred to DDM
by immobilized-metal affinity
chromatography and from DDM to
nanodiscs
Damian et al. (2012) and Casiraghi
et al. (2016)
A8-35
SMALP
The ghrelin GHS-R1a receptor was
expressed in E. coli as inclusion bodies,
solubilized in SDS, folded in A8-35,
transferred to DDM by immobilizedmetal affinity chromatography,
reconstituted into proteoliposomes, and
the proteoliposomes solubilized with
SMA
Logez et al. (2016)
A8-35, A8-75,
A34-35,
A34-75,
SAPol, NAPol
Vacuum
APol-trapped MPs were transferred to
vacuum for mass spectrometry
(MS) either by electrospray ionization
(ESI) or matrix-assisted laser desorption/
ionization (MALDI). See Chap. 14
Catoire et al. (2009), Bechara et al.
(2012), Leney et al. (2012), Hopper
et al. (2013), Calabrese et al. (2015),
and Watkinson et al. (2015, 2017)
5.7 Transferring Membrane Proteins from Amphipols to Other Environments
307
environments.
Amphipol
Final
environment Comments
References
A8-35
Another
APol
An examination of the rate of exchange
of MP-adsorbed APols when exposed to
an excess of a competing APol. In the
case of A8-35, the rate of exchange is
strongly dependent on the ionic strength
Zoonens et al. (2007)
A8-35, A8-75,
OAPA-20,
PMAL-B
Detergent or
mixed
detergent/
lipid micelles
APols can be efficiently and very rapidly
displaced from the TM surface of MPs
by an excess of detergent (cf. Fig. 5.40)
Tribet et al. (1997, 2009), Nagy et al.
(2001), and Zoonens et al. (2007)
OAPA-20,
PMAL-B,
A8-35,
NAPols
Lipid
vesicles
Transfer was initiated by directly
exposing protein/APol complexes to
preformed lipid vesicles. The first study
deals with an α-helical MP,
diacylglycerol kinase (DAGK), the
second one with the pore-forming
domain of diphtheria toxin
Nagy et al. (2001) and Kyrychenko
et al. (2012)
A8-35
Black lipid
film
The functionality of OmpA and FomA
following folding in A8-35 was
examined by applying the complexes to
black lipid films. It was noted that
recovery of native-like pore conductance
requires that the APol be applied to both
the cis and trans sides of the film
(cf. Fig. 5.35)
Pocanschi et al. (2006b)
A8-35
Live cells
A rhodamine-labeled peptide mimicking
the transmembrane anchor of neuropilin1 was trapped with NBD-labeled A8-35
and the distribution of the two species
following delivery to COS-7 cells
followed by confocal fluorescence
microscopy. See Chap. 15, Fig. 15.23
Popot et al. (2011)
A8-35
Lipidic
mesophase
Direct transfer of BR from A8-35 to a
lipidic mesophase led to the formation of
highly ordered BR crystals containing
no APol. See Chap. 11, § 11.3.2
Polovinkin et al. (2014b)
SMA
Lipidic
mesophase
Same as above, using SMALP-trapped
bacteriorhodopsin from Haloquadratum
walsbyi
Broecker et al. (2017)
A8-35
Nanodiscs
Two GPCRs were expressed in E. coli as
inclusion bodies, solubilized in SDS,
folded in A8-35, and transferred to DDM
by immobilized-metal affinity
chromatography and from DDM to
nanodiscs
Damian et al. (2012) and Casiraghi
et al. (2016)
A8-35
SMALP
The ghrelin GHS-R1a receptor was
expressed in E. coli as inclusion bodies,
solubilized in SDS, folded in A8-35,
transferred to DDM by immobilizedmetal affinity chromatography,
reconstituted into proteoliposomes, and
the proteoliposomes solubilized with
SMA
Logez et al. (2016)
A8-35, A8-75,
A34-35,
A34-75,
SAPol, NAPol
Vacuum
APol-trapped MPs were transferred to
vacuum for mass spectrometry
(MS) either by electrospray ionization
(ESI) or matrix-assisted laser desorption/
ionization (MALDI). See Chap. 14
Catoire et al. (2009), Bechara et al.
(2012), Leney et al. (2012), Hopper
et al. (2013), Calabrese et al. (2015),
and Watkinson et al. (2015, 2017)
5.7 Transferring Membrane Proteins from Amphipols to Other Environments
307