(F. Giusti, unpublished data), which ought to considerably increase the fraction of MP/APol complexes
that adsorb onto silver or gold beads (cf. Henglein and Meisel 1998) and thereby find themselves in an
appropriate position for SERS to occur.
Using labeled APols for topological or conformational studies of MPs is only in its infancy.
Study 8.26 has opened the way. Trapping a MP that carries tryptophan – or, better, 5-fluorotryptophan
(5-F-Trp), whose fluorescence decay is monoexponential – residues at single positions in its sequence
using APols that can quench their fluorescence ought to permit mapping the relative distance of each
position to the APol layer associated with the TM surface. The approach could be refined in many
different ways. Trapping a given set of mutants with a series of APols that can quench the fluorescence
from variable distances would be a way to improve distance estimates. This could be achieved by
comparing the level of quenching achieved by fluorescent APols whose absorbance bands overlap
more or less with the emission band of Trp or 5-F-Trp, thus modulating R 0 , the distance at which 50%
energy transfer takes place. A similar use could be made of a recently developed spin-labeled APol
(F. Giusti, unpublished data) or, possibly, of APols carrying bromine atoms in either their hydrophobic
or their polar head regions.
The same approach could conceivably be used to identify conformational transitions during the
functional cycle of MPs and study their kinetics.
References
Bazzacco, P., Billon-Denis, E., Sharma, K.S., Catoire, L.J., Mary, S., Le Bon, C., Point, E., Banères, J.-L., Durand, G.,
Zito, F., Pucci, B., Popot, J.-L. (2012) Non-ionic homopolymeric amphipols: Application to membrane protein
folding, cell-free synthesis, and solution NMR. Biochemistry 51:1416–1430.
Cantor, C.R., Schimmel, P.R. (1980) Biophysical Chemistry. Part II: Techniques for the study of biological structure and
function. W.H. Freeman and company, San Francisco, 846 p.
Catoire, L.J., Damian, M., Giusti, F., Martin, A., van Heijenoort, C., Popot, J.-L., Guittet, E., Banères, J.-L. (2010)
Structure of a GPCR ligand in its receptor-bound state: leukotriene B 4 adopts a highly constrained conformation when
associated to human BLT2. J. Am. Chem. Soc. 132:9049–9057.
Charvolin, D., Perez, J.-B., Rouvière, F., Giusti, F., Bazzacco, P., Abdine, A., Rappaport, F., Martinez, K.L., Popot, J.-L.
(2009) The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports.
Proc. Natl. Acad. Sci. USA 106:405–410.
Charvolin, D., Picard, M., Huang, L.-S., Berry, E.A., Popot, J.-L. (2014) Solution behavior and crystallization of
cytochrome bc 1 in the presence of amphipols. J. Membr. Biol. 247:981–996.
Craig, E.A., Gambill, B.D., Nelson, R.J. (1993) Heat shock proteins: molecular chaperones of protein biogenesis.
Microbiol. Rev. 57:402–414.
Cuevas Arenas, R., Danielczak, B., Martel, A., Porcar, L., Breyton, C., Ebel, C., Keller, S. (2017) Fast collisional lipid
transfer among polymer-bounded nanodiscs. Sci. Rep. 7:45875.
Dahmane, T. (2007) Protéines membranaires et amphipols : stabilisation, fonction, renaturation, et développement
d'amphipols sulfonatés pour la RMN des solutions. Thèse de Doctorat, Université Paris-7, Paris, 229 p.
Dahmane, T., Damian, M., Mary, S., Popot, J.-L., Banères, J.-L. (2009) Amphipol-assisted in vitro folding of G proteincoupled receptors. Biochemistry 48:6516–6521.
Dahmane, T., Rappaport, F., Popot, J.-L. (2013) Amphipol-assisted folding of bacteriorhodopsin in the presence and
absence of lipids. Functional consequences. Eur. Biophys. J. 42:85–101.
Della Pia, E.A., Holm, J., Lloret, N., Le Bon, C., Popot, J.-L., Zoonens, M., Nygård, J., Martinez, K.L. (2014a) A step
closer to membrane protein multiplexed nano-arrays using biotin-doped polypyrrole. ACS Nano 8:1844–1853.
Della Pia, E.A., Westh Hansen, R., Zoonens, M., Martinez, K.L. (2014b) Functionalized amphipols: A versatile toolbox
suitable for applications of membrane proteins in synthetic biology. J. Membr. Biol. 247:815–826.
Duarte, A.M.S., Wolfs, C.J.A.M., Koehorsta, R.B.M., Popot, J.-L., Hemminga, M.A. (2008) Solubilization of V-ATPase
transmembrane peptides by amphipol A8-35. J. Peptide Chem. 14:389–393.
Feinstein, H.E., Tifrea, D., Sun, G., Popot, J.-L., de la Maza, L.M., Cocco, M.J. (2014) Long-term stability of a vaccine
formulated with the amphipol-trapped major outer membrane protein from Chlamydia trachomatis. J. Membr. Biol.
247:1053–1065.
References
401
that adsorb onto silver or gold beads (cf. Henglein and Meisel 1998) and thereby find themselves in an
appropriate position for SERS to occur.
Using labeled APols for topological or conformational studies of MPs is only in its infancy.
Study 8.26 has opened the way. Trapping a MP that carries tryptophan – or, better, 5-fluorotryptophan
(5-F-Trp), whose fluorescence decay is monoexponential – residues at single positions in its sequence
using APols that can quench their fluorescence ought to permit mapping the relative distance of each
position to the APol layer associated with the TM surface. The approach could be refined in many
different ways. Trapping a given set of mutants with a series of APols that can quench the fluorescence
from variable distances would be a way to improve distance estimates. This could be achieved by
comparing the level of quenching achieved by fluorescent APols whose absorbance bands overlap
more or less with the emission band of Trp or 5-F-Trp, thus modulating R 0 , the distance at which 50%
energy transfer takes place. A similar use could be made of a recently developed spin-labeled APol
(F. Giusti, unpublished data) or, possibly, of APols carrying bromine atoms in either their hydrophobic
or their polar head regions.
The same approach could conceivably be used to identify conformational transitions during the
functional cycle of MPs and study their kinetics.
References
Bazzacco, P., Billon-Denis, E., Sharma, K.S., Catoire, L.J., Mary, S., Le Bon, C., Point, E., Banères, J.-L., Durand, G.,
Zito, F., Pucci, B., Popot, J.-L. (2012) Non-ionic homopolymeric amphipols: Application to membrane protein
folding, cell-free synthesis, and solution NMR. Biochemistry 51:1416–1430.
Cantor, C.R., Schimmel, P.R. (1980) Biophysical Chemistry. Part II: Techniques for the study of biological structure and
function. W.H. Freeman and company, San Francisco, 846 p.
Catoire, L.J., Damian, M., Giusti, F., Martin, A., van Heijenoort, C., Popot, J.-L., Guittet, E., Banères, J.-L. (2010)
Structure of a GPCR ligand in its receptor-bound state: leukotriene B 4 adopts a highly constrained conformation when
associated to human BLT2. J. Am. Chem. Soc. 132:9049–9057.
Charvolin, D., Perez, J.-B., Rouvière, F., Giusti, F., Bazzacco, P., Abdine, A., Rappaport, F., Martinez, K.L., Popot, J.-L.
(2009) The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports.
Proc. Natl. Acad. Sci. USA 106:405–410.
Charvolin, D., Picard, M., Huang, L.-S., Berry, E.A., Popot, J.-L. (2014) Solution behavior and crystallization of
cytochrome bc 1 in the presence of amphipols. J. Membr. Biol. 247:981–996.
Craig, E.A., Gambill, B.D., Nelson, R.J. (1993) Heat shock proteins: molecular chaperones of protein biogenesis.
Microbiol. Rev. 57:402–414.
Cuevas Arenas, R., Danielczak, B., Martel, A., Porcar, L., Breyton, C., Ebel, C., Keller, S. (2017) Fast collisional lipid
transfer among polymer-bounded nanodiscs. Sci. Rep. 7:45875.
Dahmane, T. (2007) Protéines membranaires et amphipols : stabilisation, fonction, renaturation, et développement
d'amphipols sulfonatés pour la RMN des solutions. Thèse de Doctorat, Université Paris-7, Paris, 229 p.
Dahmane, T., Damian, M., Mary, S., Popot, J.-L., Banères, J.-L. (2009) Amphipol-assisted in vitro folding of G proteincoupled receptors. Biochemistry 48:6516–6521.
Dahmane, T., Rappaport, F., Popot, J.-L. (2013) Amphipol-assisted folding of bacteriorhodopsin in the presence and
absence of lipids. Functional consequences. Eur. Biophys. J. 42:85–101.
Della Pia, E.A., Holm, J., Lloret, N., Le Bon, C., Popot, J.-L., Zoonens, M., Nygård, J., Martinez, K.L. (2014a) A step
closer to membrane protein multiplexed nano-arrays using biotin-doped polypyrrole. ACS Nano 8:1844–1853.
Della Pia, E.A., Westh Hansen, R., Zoonens, M., Martinez, K.L. (2014b) Functionalized amphipols: A versatile toolbox
suitable for applications of membrane proteins in synthetic biology. J. Membr. Biol. 247:815–826.
Duarte, A.M.S., Wolfs, C.J.A.M., Koehorsta, R.B.M., Popot, J.-L., Hemminga, M.A. (2008) Solubilization of V-ATPase
transmembrane peptides by amphipol A8-35. J. Peptide Chem. 14:389–393.
Feinstein, H.E., Tifrea, D., Sun, G., Popot, J.-L., de la Maza, L.M., Cocco, M.J. (2014) Long-term stability of a vaccine
formulated with the amphipol-trapped major outer membrane protein from Chlamydia trachomatis. J. Membr. Biol.
247:1053–1065.
References
401