Anomeric Effects on the Structure of Micelles of Alkyl Maltosides in Water

Dupuy, Camille; Auvray, X.; Petipas, C.; Rico‐Lattes, Isabelle; Lattes, Armand

doi:10.1021/la9604285

Cited by 115 publications

(177 citation statements)

References 9 publications

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“…These results suggest that high DM concentrations can affect the Rho oligomeric structure and slow down the Gt activation process. Our results are in agreement with a previous study where a decrease in Gt activity with increasing DM concentration was reported (47,48). This experiment is not reliable for the cross-linked Rho, since modification of the surface of the receptor alters activation of Gt (57)(58)(59).…”

Section: Oligomeric Structure Of Rho In Chaps-we Also Investigated Thsupporting

confidence: 90%

“…The molecular mass of the Rho monomer and dimer estimated by SDS-PAGE is ϳ36 and 72 kDa, respectively. 3 The aggregation number of DM is 132 (48), and the molecular mass of DM micelles is ϳ67,399 Da (48). The sizes of the Rho monomer and Rho dimer complexes with DM seen in the size exclusion chromatography experiments (ϳ90 -100 and ϳ140 -160 kDa, respectively) agree with the sums of the protein and micellar molecular masses (103,500 Da (monomer) and 149,500 Da (dimer)).…”

Section: Resultsmentioning

confidence: 58%

“…However, another explanation supported by experiments is also possible. At least in water, DM micelles are highly elliptical, with a shorter radius, 0.59 of the length of the longer radius, and contain n ϭ 132 detergent molecules (48). Rho probably localizes to portions of the ellipsoid with less curvature (i.e.…”

Section: Oligomeric Structure Of Rho In Chaps-we Also Investigated Thmentioning

confidence: 99%

See 2 more Smart Citations

Functional Characterization of Rhodopsin Monomers and Dimers in Detergents

Jastrzębska

Maeda

Zhu

et al. 2004

Journal of Biological Chemistry

122

126

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Rhodopsin (Rho) is a G protein-coupled receptor that initiates phototransduction in rod photoreceptors. High expression levels of Rho in the disc membranes of rod outer segments and the propensity of Rho to form higher oligomeric structures are evident from atomic force microscopy, transmission electron microscopy, and chemical cross-linking experiments. To explore the structural and functional properties of Rho in n-dodecyl-␤-maltoside, frequently used to purify heterologously expressed Rho and its mutants, we used gel filtration techniques, blue native gel electrophoresis, and functional assays. Here, we show that in micelles containing n-dodecyl-␤-maltoside at concentrations greater than 3 mM, Rho is present as a single monomer per detergent micelle. In contrast, in 12 mM 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propanesulfonate (CHAPS), micelles contain mostly dimeric Rho. The cognate G protein transducin (Gt) appears to have a preference for binding to the Rho dimer, and the complexes fall apart in the presence of guanosine 5-3-O-(thio)triphosphate. Cross-linked Rho dimers release the chromophore at a slower rate than monomers and are much more resistant to heat denaturation. Both Rho* monomers and dimers are capable of activating Gt, and both of them are phosphorylated by Rho kinase. Rho expressed in HEK293 cells is also readily cross-linked by a bifunctional reagent. These studies provide an explanation of how detergent influences the oligomer-dimermonomer equilibrium of Rho and describe the functional characterization of Rho monomers and dimers in detergent.Rhodopsin (Rho), 1 the prototypical G protein-coupled receptor (GPCR), functions in the absorption of a photon in retinal rod photoreceptors (1-3). As are other GPCRs (4, 5), Rho is a seven-transmembrane-spanning helical protein (6). The high expression level of Rho, its specific localization in the internal discs of the structures termed rod outer segments (ROS), and the lack of other highly abundant membrane proteins allow Rho to be imaged in the native disc membranes by atomic force microscopy and transmission electron microscopy (7-9). These images have revealed rows of Rho dimers in native disc membranes. Subsequently, BN-and SDS-PAGE, chemical crosslinking, and proteolysis experiments corroborated that Rho consists mainly of dimers and higher oligomers in disc membranes.2 Medina et al. (7) reported that Rho and photoactivated Rho (Rho*) preserved a dimeric quaternary structure in detergent.These results are in conflict with a model of rapidly diffusing Rho in the "mosaic" fluid disc membrane (10) supported by measurements of Rho diffusion and rotation in disc membranes and by low resolution neutron diffraction studies (11)(12)(13)(14). The concept of rapidly diffusing monomeric molecules is at variance with the dimeric forms of other GPCRs (15,16). Moreover, the size of the G protein and arrestin surface interacting with Rho is almost twice as large as the exposed cytoplasmic surface of a single Rho molecule (17,18).Nonphysiological dimers of R...

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Section: Oligomeric Structure Of Rho In Chaps-we Also Investigated Thsupporting

confidence: 90%

Section: Resultsmentioning

confidence: 58%

Section: Oligomeric Structure Of Rho In Chaps-we Also Investigated Thmentioning

confidence: 99%

See 1 more Smart Citation

Functional Characterization of Rhodopsin Monomers and Dimers in Detergents

Jastrzębska

Maeda

Zhu

et al. 2004

Journal of Biological Chemistry

122

126

View full text Add to dashboard Cite

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“…We use their values for C 10 G 2 and C 12 G 2 (ESI, † Table 5) together with an interpolated value for C 11 G 2 . The values are in good agreement with data from independent sources 47,48 (see also ref. 8).…”

Section: Determination Of Cmcsupporting

confidence: 89%

The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization

Müh

DiFiore

Zouni

2015

Phys. Chem. Chem. Phys.

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With the aim of better understanding the phase behavior of alkyl maltosides (n-alkyl-b-D-maltosides, C n G 2 ) under the conditions of membrane protein crystallization, we studied the influence of poly(ethylene glycol) (PEG) 2000, a commonly used precipitating agent, on the critical micelle concentration (CMC) of the alkyl maltosides by systematic variation of the number n of carbon atoms in the alkyl chain (n = 10, 11, and 12) and the concentration of PEG2000 (w) in a buffer suitable for the crystallization of cyanobacterial photosystem II. CMC measurements were based on established fluorescence techniques using pyrene and 8-anilinonaphthalene-1-sulfonate (ANS). We found an increase of the CMC with increasing PEG concentration according to ln(CMC/CMC 0 ) = k P w, where CMC 0 is the CMC in the absence of PEG and k P is a constant that we termed the ''polymer constant''. In parallel, we measured the influence of PEG2000 on the surface tension of detergent-free buffer solutions. At PEG concentrations w 4 1% w/v, the surface pressure p s (w) = g(0) À g(w) was found to depend linearly on the PEG concentration according to p s (w) = kw + p s (0), where g (0) is the surface tension in the absence of PEG. Based on a molecular thermodynamic modeling, CMC shifts and surface pressure due to PEG are related, and it is shown that k P = kc(n) + Z, where c(n) is a detergent-specific constant depending inter alia on the alkyl chain length n and Z is a correction for molarity. Thus, knowledge of the surface pressure in the absence of a detergent allows for the prediction of the CMC shift. The PEG effect on the CMC is discussed concerning its molecular origin and its implications for membrane protein solubilization and crystallization.

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“…5a). In fact, a study by Dupuy et al (1997) indicates that -maltoside molecules form oblate ellipsoidal micelles in water. Nevertheless, an attempt to simulate the detergent-solubilized truncated KcsA K + channel was able to essentially reproduce the low-angle scattering features including the characteristic minimum near q = 0.11 Å À1 , where q ¼ ð4=Þ sin , 2 is the scattering angle and is the wavelength (Fig.…”

Section: Figurementioning

confidence: 99%

Biological solution scattering: recent achievements and future challenges

Grossmann

2007

J Appl Cryst

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In the post-genomic age it is apparent that as structures of larger macromolecules and their complexes are investigated, structure-function investigations are often confronted with the necessity to apply a portfolio of tools for biochemical and biophysical characterization. A survey of the published literature over the last decade reveals that publications in the area of structural biology employing neutron or X-ray scattering as one of their techniques tripled since 1995. Yet, taken as a whole, the contribution from smallangle scattering (SAS) to research papers dealing with structural analyses is still only of the order of 1% in 2005 (for comparison, less than 0.5% in 1995). Nevertheless, the last few years saw stimulating biological applications and analysis procedures which emphasize the growing potential of SAS applications for the structural studies of macromolecules in solution. The usage of SAS largely consists of low-resolution reconstructions of molecules with partial or without presumption of structural details, consistency analysis of high-resolution crystallographic structures and their corresponding low-resolution models determined in the solution state, and rigid-body refinement of multi-subunit assemblies including complexes and full-length multidomain proteins. Complementary structural information obtained from SAS in conjunction with data acquired by protein crystallography, NMR, molecular dynamics or computational docking provides a means to link low-and high-resolution models essential for the elucidation of biomolecular organization, interactions and function. The capabilities as well as limitations of determining low-resolution structures of multidomain proteins, macromolecular complexes and assemblies are highlighted in three examples. The first example is the characterization of the conformation of the PDZ region of SAP97, a multidomain protein involved in the regulation and localization of membrane receptor molecules. The second example is the characterization of the structural features of the TIM10 complex, an escort molecule for mitochondrial inner-membrane proteins, and the third example is the description of the shape and pH-induced conformational transition of a full-length bacterial potassium channel. The latter two in particular benefited from neutron scattering with contrast variation by using H-D labelling of the macromolecular complex or the solvent, respectively.

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Anomeric Effects on the Structure of Micelles of Alkyl Maltosides in Water

Cited by 115 publications

References 9 publications

Functional Characterization of Rhodopsin Monomers and Dimers in Detergents

Functional Characterization of Rhodopsin Monomers and Dimers in Detergents

The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization

Biological solution scattering: recent achievements and future challenges

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