TY - JOUR
T1 - NMR investigation of the multidrug transporter EmrE, an integral membrane protein
AU - Schwaiger, Manfred
AU - Lebendiker, Mario
AU - Yerushalmi, Hagit
AU - Coles, Murray
AU - Gröger, Adriane
AU - Schwarz, Christian
AU - Schuldiner, Shimon
AU - Kessler, Horst
PY - 1998/6/15
Y1 - 1998/6/15
N2 - EmrE is an Escherichia coli multidrug transport protein that confers resistance to a wide range of toxicants by active transport across the bacterial cell membrane. The highly hydrophobic polytopic integral membrane protein has been purified and studied in its full-length form by high- resolution NMR spectroscopy in a mixture of chloroform/methanol/water (6:6: 1, by vol.). Full activity is maintained after reconstitution of the protein into proteoliposomes from this solvent mixture. A series of heteronuclear (1H-15N) two- and three-dimensional experiments, as well as triple resonance experiments, were applied to the 110-residue protein and led to the assignment of the 1H, 15N and a large part of the 13C backbone resonances as well as many of the sidechain resonances. A preliminary analysis of the secondary structure, based on sequential NOE connectivities, deviation of chemical shifts from random coil values and 3J(NH-Hα) coupling constants supports a model where the protein forms four α-helices between residues 4-26 (TM1), 32-53 (TM2), 58-76 (TM3) and 85-106 (TM4). For the residues of helices TM2 and TM3 a significant line broadening occurs due to slow conformational processes.
AB - EmrE is an Escherichia coli multidrug transport protein that confers resistance to a wide range of toxicants by active transport across the bacterial cell membrane. The highly hydrophobic polytopic integral membrane protein has been purified and studied in its full-length form by high- resolution NMR spectroscopy in a mixture of chloroform/methanol/water (6:6: 1, by vol.). Full activity is maintained after reconstitution of the protein into proteoliposomes from this solvent mixture. A series of heteronuclear (1H-15N) two- and three-dimensional experiments, as well as triple resonance experiments, were applied to the 110-residue protein and led to the assignment of the 1H, 15N and a large part of the 13C backbone resonances as well as many of the sidechain resonances. A preliminary analysis of the secondary structure, based on sequential NOE connectivities, deviation of chemical shifts from random coil values and 3J(NH-Hα) coupling constants supports a model where the protein forms four α-helices between residues 4-26 (TM1), 32-53 (TM2), 58-76 (TM3) and 85-106 (TM4). For the residues of helices TM2 and TM3 a significant line broadening occurs due to slow conformational processes.
KW - EmrE protein
KW - Membrane protein
KW - Multidrug resistance
KW - NMR
KW - Secondary structure
UR - http://www.scopus.com/inward/record.url?scp=0032526126&partnerID=8YFLogxK
U2 - 10.1046/j.1432-1327.1998.2540610.x
DO - 10.1046/j.1432-1327.1998.2540610.x
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C2 - 9688273
AN - SCOPUS:0032526126
SN - 0014-2956
VL - 254
SP - 610
EP - 619
JO - European Journal of Biochemistry
JF - European Journal of Biochemistry
IS - 3
ER -