title: Synthesis of uniformly deuterated n-dodecyl-β-D-maltoside (d39-DDM) for solubilization of membrane proteins in TROSY NMR experiments.
abstract: This work reports the first synthesis of uniformly deuterated n-dodecyl-β-D-maltoside (d39-DDM). DDM is a mild non-ionic detergent often used in the extraction and purification of membrane proteins and for solubilizing them in experimental studies of their structure, dynamics and binding of ligands. We required d39-DDM for solubilizing large α-helical membrane proteins in samples for [(15)N-(1)H]TROSY (transverse relaxation-optimized spectroscopy) NMR experiments to achieve the highest sensitivity and best resolved spectra possible. Our synthesis of d39-DDM used d7-D-glucose and d25-n-dodecanol to introduce deuterium labelling into both the maltoside and dodecyl moieties, respectively. Two glucose molecules, one converted to a glycosyl acceptor with a free C4 hydroxyl group and one converted to a glycosyl donor substituted at C1 with a bromine in the α-configuration, were coupled together with an α(1 → 4) glycosidic bond to give maltose, which was then coupled with n-dodecanol by its substitution of a C1 bromine in the α-configuration to give DDM. (1)H NMR spectra were used to confirm a high level of deuteration in the synthesized d39-DDM and to demonstrate its use in eliminating interfering signals from TROSY NMR spectra of a 52-kDa sugar transport protein solubilized in DDM.
authors.affiliation: School of Molecular and Cellular Biology, University of Leeds, Leeds, UK; Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK; Waseda University, Tokyo, Japan.
authors.forename: Kazumi
authors.initials: K
authors.lastname: Hiruma-Shimizu
authors.forename: Arnout P
authors.initials: AP
authors.lastname: Kalverda
authors.forename: Peter J F
authors.initials: PJ
authors.lastname: Henderson
authors.forename: Steve W
authors.initials: SW
authors.lastname: Homans
authors.forename: Simon G
authors.initials: SG
authors.lastname: Patching
mesh_terms: D002135:Calcium-Binding Proteins / Q000737:chemistry*; D060326:Chemistry Techniques, Synthetic; D003902:Detergents / Q000138:chemical synthesis* / Q000737:chemistry*; D003903:Deuterium / Q000737:chemistry*; D005960:Glucosides / Q000138:chemical synthesis* / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000737:chemistry*; D033902:Periplasmic Binding Proteins / Q000737:chemistry*; D012995:Solubility
chemical_list: D002135:Calcium-Binding Proteins; D003902:Detergents; D005960:Glucosides; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; C023550:galactose-binding protein; C040358:dodecyl maltoside; D003903:Deuterium
keywords: NMR; detergent; deuteration; membrane proteins; n-dodecyl-β-d-maltoside