Purification of octyl β-d-glucopyranoside and re-estimation of its micellar size

Lorber, Bernard; Bishop, John Bradford; DeLucas, Lawrence J.

doi:10.1016/0005-2736(90)90421-j

Cited by 85 publications

(74 citation statements)

References 26 publications

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“…Similar measurement revealed a range of molecular weight from 25 000 to 36 000, depending on the temperature, for the OG micelles at low concentration (∼1%) (19). The molecular weights were also confirmed by a combination of sedimentation velocity and diffusion experiments (18). A somewhat smaller value of R g ) 17.7 Å was obtained for 1%(w/v) OG aqueous solution from another SANS measurement (20).…”

Section: Estimation Of the Micelle Sizes Of Og Alone At High Concentrsupporting

confidence: 56%

“…The sizes of OG micelles determined from this study are compatible with those previously reported using other techniques. Dynamic light scattering yielded hydrodynamic radii of 23 ( 3, 28 ( 3, and 31 ( 3 Å for the OG micelles at concentration of 1, 2, and 5%(w/v), respectively (18). Similar measurement revealed a range of molecular weight from 25 000 to 36 000, depending on the temperature, for the OG micelles at low concentration (∼1%) (19).…”

Section: Estimation Of the Micelle Sizes Of Og Alone At High Concentrmentioning

confidence: 67%

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Determination of the B820 Subunit Size of a Bacterial Core Light-Harvesting Complex by Small-Angle Neutron Scattering

et al. 2003

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The B820 subunit is an integral pigment-membrane protein complex and can be obtained by both dissociation of the core light-harvesting complex (LH1) in photosynthetic bacteria and reconstitution from its component parts in the presence of n-octyl -D-glucopyranoside (OG). Intrinsic size of the B820 subunit from Rhodospirillum rubrum LH1 complex was measured by small-angle neutron scattering in perdeuterated OG solution and evaluated by Guinier analysis. Both the B820 subunits prepared by dissociation of LH1 and reconstitution from apopolypeptides and pigments were shown to have a molecular weight of 11 400 ( 500 and radius of gyration of 11.0 ( 1.0 Å, corresponding to a heterodimer consisting of one pair of R -polypeptides and two bacteriochlorophyll a molecules. Molecular weights of micelles formed by OG alone in solutions were determined in a range from 30 000 to 50 000 over concentrations of 1-5% (w/v), and thus are much larger than that of the B820 subunit. Similar measurement on the pigment-depleted apopolypeptides revealed highly heterogeneous behavior in the OG solutions, indicating that aggregates with various sizes were formed. The result provides evidence that bacteriochlorophyll a molecules play a crucial role in stabilizing and maintaining the B820 subunits in the dimeric state in solution. Further measurements on individual R-and -polypeptides exhibited a marked difference in aggregation property between the two polypeptides. The R-polypeptides appear to be uniformly dissolved in OG solution in a monomeric form, whereas the -polypeptides favor a self-associated form and tend to form large aggregates even in the presence of detergent. The difference in aggregation tendency was discussed in relation to the different behavior between R-and -polypeptides in reconstitution with bacteriochlorophyll a molecules.Despite intense interest and continuing investigation, controversy over the nature of an intermediate form of bacterial core light-harvesting complexes (LH1 1 ) has not been settled. The LH1 complex, serving as highly efficient molecular machinery for collection and transfer of solar energy to photochemical reaction centers in photosynthetic bacteria, is a large multimer of a minimal unit composed of two small integral membrane polypeptides (R and , ca. 6 kDa) associated with two bacteriochlorophyll (BChl) and one carotenoid molecule. Upon addition of detergents, the LH1 multimer can be dissociated into a structural subunit characterized by a Q y absorption band at 820 nm (1). Both the subunit (referred to as B820) and LH1 complex can be reversibly dissociated and reconstituted from the their components (2). Early chromatographic studies by gel filtration estimated a range of molecular weight from 32 to 65 kDa for the B820 subunit, roughly corresponding to a tetramer composed of two minimal structural units (1, 3). In contrast, spectroscopic evidence suggested that the B820 is a heterodimer consisting of two interacting BChl a molecules linked to one R -polypeptide pair (4,5). Recently, ide...

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Section: Estimation Of the Micelle Sizes Of Og Alone At High Concentrsupporting

confidence: 56%

Section: Estimation Of the Micelle Sizes Of Og Alone At High Concentrmentioning

confidence: 67%

Determination of the B820 Subunit Size of a Bacterial Core Light-Harvesting Complex by Small-Angle Neutron Scattering

et al. 2003

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“…We assumed that p250 and p160 associate with one detergent micelle each and that M␣ϩM␤Ј and M␣ do not associate with detergent at all. Octylglucoside micelle size was estimated to be 25 kDa (45). The partial specific volume of each species was estimated by summing the partial specific volumes of each amino acid, the N-linked glycan of the high mannose or complex type, 20 kDa of O-linked glycans for each RPT domain, and the octylglucoside detergent micelle for those species in which it was warranted (43).…”

Section: Molecular Weight Calculationsmentioning

confidence: 99%

The Kringle-like Domain Facilitates Post-endoplasmic Reticulum Changes to Premelanosome Protein (PMEL) Oligomerization and Disulfide Bond Configuration and Promotes Amyloid Formation

Watt

Spruce

et al. 2016

Journal of Biological Chemistry

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The formation of functional amyloid must be carefully regulated to prevent the accumulation of potentially toxic products. Premelanosome protein (PMEL) forms non-toxic functional amyloid fibrils that assemble into sheets upon which melanins ultimately are deposited within the melanosomes of pigment cells. PMEL is synthesized in the endoplasmic reticulum but forms amyloid only within post-Golgi melanosome precursors; thus, PMEL must traverse the secretory pathway in a non-amyloid form. Here, we identified two pre-amyloid PMEL intermediates that likely regulate the timing of fibril formation. Analyses by non-reducing SDS-PAGE, size exclusion chromatography, and sedimentation velocity revealed two native high M r disulfide-bonded species that contain Golgi-modified forms of PMEL. These species correspond to disulfide bond-containing dimeric and monomeric PMEL isoforms that contain no other proteins as judged by two-dimensional PAGE of metabolically labeled/immunoprecipitated PMEL and by mass spectrometry of affinity-purified complexes. Metabolic pulse-chase analyses, small molecule inhibitor treatments, and evaluation of site-directed mutants suggest that the PMEL dimer forms around the time of endoplasmic reticulum exit and is resolved by disulfide bond rearrangement into a monomeric form within the late Golgi or a post-Golgi compartment. Mutagenesis of individual cysteine residues within the non-amyloid cysteine-rich Kringlelike domain stabilizes the disulfide-bonded dimer and impairs fibril formation as determined by electron microscopy. Our data show that the Kringle-like domain facilitates the resolution of disulfide-bonded PMEL dimers and promotes PMEL functional amyloid formation, thereby suggesting that PMEL dimers must be resolved to monomers to generate functional amyloid fibrils.

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“…It has one sugar moiety less in its head group compared to maltosides ( Figure 5), but similarly to them, OG is a non-ionic, albeit less milder detergent. It has a very high CMC of 0.53%/20 mM [53] with the compact micelle (Mw~25 kDa [53] …”

Section: N-octyl-β-d-glucopyranosidementioning

confidence: 99%

An Overview of the Top Ten Detergents Used for Membrane Protein Crystallization

2017

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Abstract:To study integral membrane proteins, one has to extract them from the membrane-the step that is typically achieved by the application of detergents. In this mini-review, we summarize the top 10 detergents used for the structural analysis of membrane proteins based on the published results. The aim of this study is to provide the reader with an overview of the main properties of available detergents (critical micelle concentration (CMC) value, micelle size, etc.) and provide an idea of what detergents to may merit further study. Furthermore, we briefly discuss alternative solubilization and stabilization agents, such as polymers.

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Purification of octyl β-d-glucopyranoside and re-estimation of its micellar size

Cited by 85 publications

References 26 publications

Determination of the B820 Subunit Size of a Bacterial Core Light-Harvesting Complex by Small-Angle Neutron Scattering

Determination of the B820 Subunit Size of a Bacterial Core Light-Harvesting Complex by Small-Angle Neutron Scattering

The Kringle-like Domain Facilitates Post-endoplasmic Reticulum Changes to Premelanosome Protein (PMEL) Oligomerization and Disulfide Bond Configuration and Promotes Amyloid Formation

An Overview of the Top Ten Detergents Used for Membrane Protein Crystallization

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