Location and orientation relative to the micelle surface for glucagon in mixed micelles with dodecylphosphocholine EPR and NMR studies

Brown, L. R.; Bösch, Chris; Wüthrich, Kurt

doi:10.1016/0005-2736(81)90447-8

Cited by 126 publications

(134 citation statements)

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“…Llinas et al (1978) calculated the T, of alumichrome, a metal binding cyclohexapeptide, of -9.4 (-loglo(T, sec)) from backbone amide proton relaxation rate data in DMSO. Brown (1981) calculated the correlation times of 5-doxylstearic acid in solution and in a glucagon/DPC micelle complex as 9.9 and 8.8 (-loglo(T, sec)), respectively, from EPR hyperfine splitting constants. Yuan et al (1996) calculated an overall correlation time of 8 * 0.1 (-logIo(T, sec)) for melittin bound to MMPC micelles from I3C-NMR relaxation measurements.…”

Section: Correlation Time Calculationsmentioning

confidence: 99%

“…surface and for comparison to the amide exchange and relaxation time measurement data. Other studies have established experimental conditions for the use of spin-labeled lipids in DPC and SDS micelles to evaluate peptide orientation at lipid surfaces (Brown, 1981;Van den Hooven et al, 1996). Similar to these studies, in this work a ratio of peptide to SDS monomer to 5-doxylstearic acid of 1 :SO: 1 was used to determine the orientation of the peptide at the micelle by observation of selective line-broadening of SP protons in 2D TOCSY spectra.…”

Section: Spin Labeling Experimentsmentioning

confidence: 99%

“…Previous work has established that the 5-doxy1 moiety of the lipid 5-doxy1 stearic acid is located close to the surface of both SDS and DPC micelles (Brown, 1981;Mendz et al, 1988Mendz et al, , 1991Papavoine et al, 1994;Van den Hooven et al, 1996). The unpaired electron of the spin label broadens protons within -15 A sphere via electron-dipole nuclear-dipole interactions.…”

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confidence: 97%

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The orientation and dynamics of substance P in lipid environments

Keire

Kobayashi

1998

Protein Science

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The membrane-associated conformation of substance P (RPKPQQFFGLM-NH2) has been previously proposed to be the NKI-receptor-active conformation. In this work, NMR methods are applied to explore the orientation and dynamics of substance P at lipid surfaces for which the peptide's three-dimensional structure had been previously determined. Here the presence of dodecylphosphocholine (DPC) or sodium dodecylsulfate (SDS) micelles has been found to cause sequence specific changes in the acid-and base-catalyzed amide proton exchange rates relative to the solution state values. On binding of substance P to SDS micelles, the FFG portion showed the largest decreases in the base-catalyzed amide exchange rates. Similar sequence-specific changes in substance P are observed in the presence of DPC micelles, albeit at much weaker levels due to fast exchange between free and bound forms of the peptide. These differences are attributed to the location of the amide protons either in the surface double layer (via electrostatic effect) or inserted into the polar head group region of the micelles (via low dielectric). The sequence-specific effects of micelle association were also observed in the homonuclear nonselective spin-lattice relaxation time; these, in combination with spin-spin relaxation times, were used to calculate correlation times for the backbone amide protons. These data combined with paramagnetic broadening observations on peptide protons in the presence of spin-labeled lipids yield a detailed model of the interaction of substance P with lipid surfaces.Keywords: amide exchange; correlation times; dodecylphosphocholine; NMR; peptide-lipid interactions; sodium dodecylsulfate; substance P; TI and T2 relaxation Substance P (SP) is a member of the tachykinin family of neuropeptides and is widely distributed in both the central and peripheral nervous system of humans. Tachykinins, which are characterized by a common C-terminal tail (F-x-GLM-NH2 where x is F or V for mammals), are of biomedical interest because they have been shown to cause hypotension, contraction of smooth muscles, salivation, transmission of pain, and inflammation (Pernow, 1983). Ligands of SP receptors have potential as a novel class of anti-inflammatory or pain medication, and as a result, interest is high in the structural elements of the peptide that may control receptor activity and selectivity.The membrane-bound conformation of SP has been proposed to be the NK1 receptor active form (Schwyzer, 1995), and the three-dimensional structure of SP in the presence of Abbreviations: DPC, dodecylphosphocholine; RMSD, root-mean-square deviation; SDS, sodium dodecylsulfate; SP, substance P; 2D TOCSY, twodimensional total correlation spectroscopy. 1996). Both structures were similar, although fast exchange between free and bound forms was observed for SP with DPC micelles and predominantly bound characteristics were found for SP with SDS micelles.The three-dimensional SP structure at these lipid-membrane mimetics may represent either a receptor-bound co...

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Section: Correlation Time Calculationsmentioning

confidence: 99%

Section: Spin Labeling Experimentsmentioning

confidence: 99%

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confidence: 97%

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The orientation and dynamics of substance P in lipid environments

Keire

Kobayashi

1998

Protein Science

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“…All homonuclear and I5N-edited experiments were recorded using the time proportional phase incrementation (TPPI) method (Marion and Wuthrich, 1983). For signal accumulation in the I3C dimension, STATES-TPPI was used (States et al, 1982;Marion et al, 1989b Brown et al (1981). Hence, at the conditions used for the NMR experiments (2 mM gVIIlp, 1.5 -5 mM spin-labelled molecule, and 400 mM DodPCho) less than one molecule gVIIIp or spinlabel is bound per micelle (more than three micelles for each gVIIIp molecule).…”

Section: The Location Of the Major Coat Protein In Dodpcho Micellesmentioning

confidence: 99%

NMR Studies of the Major Coat Protein of Bacteriophage M13

Papavoine

Aelen

Konings

et al. 1995

European Journal of Biochemistry

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The membrane-bound form of the major coat protein (gV11Ip) of bacteriophage M I 3 has been studied using nuclear magnetic resonance spectroscopy. As membrane mimetics, we used dodecylphosphocholine (DodPCho) detergent micelles to solubilize the protein. We were able to nearly completely assign all resonances of the protein solubilized in DodPCho micelles by using both homonuclear and heteronuclear multidimensional experiments. Based on the patterns of the nuclear Overhauser enhancements and the chemical shifts of the resonances, we deduced the secondary structure of the protein. Additional structural information was obtained from amide proton exchange data and J-coupling constants. The protein consists of two a-helices which are connected by a hinge region around residue 21. From spin-label experiments, the location of the protein relative to the DodPCho micelles was determined. One, hydrophobic, helix spans the micelle, and another, amphipathic, helix, is located beneath the surface of the micelle. For gVTTTp in SDS micelles, we found a micellar structure which is distorted near the C-terminus of the protein; whereas for DodPCho micelles, both distorted and regular elliptical micelles occur. This distortion is probably due to the interaction of the positively charged lysine side chains with the negatively charged head group of the detergent molecules.Keywords: major coat protein; bacteriophage M I 3 ; NMR; dodecylphosphocholine; structural analysis.Our knowledge of biological processes involving membranebound proteins is limited in comparison with those involving cytoplasmic proteins. This is due, in large part, to the lack of structural information caused by difficulties in applying the methods of X-ray diffraction and NMR to membrane-bound proteins. Membrane-associated proteins are more difficult to crystallize than globular proteins. Multidimensional high-resolution NMR methods, which work well in the elucidation of the structures of small water-soluble proteins (Wiithrich, 1986) are less well-suited for studying large, slowly reorienting complexes, like membrane-bound proteins. To obtain structural information for these proteins with these techniques, model systems, mostly in the form of detergent micelles, are used as membrane mimetics. In high-resolution NMR, glucagon (Braun et al., 1981) and melittin (Brown et al., 1982) were the initial model systems Correspondence to F. J. M. Van de Ven,

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“…ESR spectra were recorded at 313 K on a Bruker ESP300 X-band spectrometer as described previously (Brown et al, 1981 ;Mctidz ct al., 1988Mctidz ct al., , 1991. The pH of the inicellar samples was 3.5 (pH meter reading).…”

Section: Chemicals Nisin Was Obtained As a Gift Of Aplin And Barrettmentioning

confidence: 99%

Surface Location and Orientation of the Lantibiotic Nisin Bound to Membrane‐Mimicking Micelles of Dodecylphosphocholine and of Sodium Dodecylsulphate

Hooven

Spronk

Kamp

et al. 1996

European Journal of Biochemistry

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The interaction of nisin, a membrane-interacting cationic polypeptide, with membranemimicking micelles of zwitterionic dodecylphosphocholine and of anionic sodium dodecylsulphate was studied. Direct contacts have been established through the observation of NOES between nisin and micelle protons. Spin-labeled DOXYL-stearic acids were incorporated into the two micellar systems. From the paramagnetic broadening effects induced in the 'H-NMR spectrum of nisin it is concluded that the molecule is localized at the surface of the micelles. Biochem. 235,. The hydrophobic residues are immersed into the micelles and oriented towards the center, whereas the more polar or charged residues have an outward orientation. The micellar systems are considered to model the first step in the mechanism of antimicrobial action of nisin, this step is the binding of nisin to the cytoplasmic membrane of target bacteria. Detailed information on this initial binding step is obtained. Hydrophobic and electrostatic interactions appear to be involved in the nisin-micelle contacts. It is suggested that subtilin, a lantibiotic structurally related to nisin, has a comparable membrane interaction surface.Keywords: bacteriocin; lanthionine-containing polypeptide; inode of action; NMR; spin labelThe polycyclic 34-residue-containing polypeptide nisin (Gross and Morel], 1971) ( Fig. 1) belongs to the class of lanthionine-containing polypeptides known as lantibiotics (Schnell et al., 1988). They are characterized by the occurrence of unusual amino acids, such as lanthionine, 3methyllanthionine, dehydroalanine (Dha) and dehydrobutyrine (Dhb). Lantibiotics are the end products of ribosomal synthesis and post-translational modifications. They are produced by a large number of Grampositive bacteria; the producer of nisin is Lactococcus lactis subsp. lactis. The post-translational modification involves dehydration of serines and threonines resulting in cx,p-unsaturated amino acids, of which some react with thiol groups of cysteine residues to form lanthionine or 3-methyllanthionine (Ingram, 1970).Nisin possesses bactericidal activity against a wide range of Gram-positive bacteria or their spores. Because of this activity it is exploited as a food preservative in the dairy and canning Correspondence fo C. W. Hilbers, NSR Center, Faculty of Science, Laboratory of Biophysical Chemistry, Toernooiveld, NL-6525 ED Nijmegen, The Netherlands Fux: +31 24 3652112. Ahhvevicztions. Ah:, 3-methylalanyl moiety of (2S, 3S, 6X)-3-metliyllanthionine; Ah,, i,-alanyl moiety of tnesn-lanthionine; .Ah, L-alanyl moicty of meso-lanthioninc or of (2S, 3S, hR)-3-methyllanthionine; 1/2/

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Location and orientation relative to the micelle surface for glucagon in mixed micelles with dodecylphosphocholine EPR and NMR studies

Cited by 126 publications

References 14 publications

The orientation and dynamics of substance P in lipid environments

The orientation and dynamics of substance P in lipid environments

NMR Studies of the Major Coat Protein of Bacteriophage M13

Surface Location and Orientation of the Lantibiotic Nisin Bound to Membrane‐Mimicking Micelles of Dodecylphosphocholine and of Sodium Dodecylsulphate

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