Heavy Atom Detergent/Lipid Combined X-ray Crystallography for Elucidating the Structure-Function Relationships of Membrane Proteins

Hanashima, Shinya; Nakane, Takanori; Mizohata, Eiichi

doi:10.3390/membranes11110823

Cited by 7 publications

(2 citation statements)

References 76 publications

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“…While Se-Met is the most popular labelling agent employed for de novo phasing, other non-amino acid Se-containing compounds have been used successfully with membrane proteins. A recent review by Hanashima et al [62] describes the application of heavy atom-labelled detergents and lipids for locating them in and for the experimental phasing of membrane protein crystal structures. Thus, for example, the common detergent dodecylβ-D-maltoside (DDM) was modified to include a selenium atom in the sugar head group and used to phase the structures of a leukotriene C4 synthase and a pentameric ligandgated ion channel.…”

Section: Discussionmentioning

confidence: 99%

Se-MAG Is a Convenient Additive for Experimental Phasing and Structure Determination of Membrane Proteins Crystallised by the Lipid Cubic Phase (In Meso) Method

Boland,

Huang,

Shanker Kaki

et al. 2023

Crystals

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Both intensity and phase information are needed for structure determination by macromolecular X-ray crystallography. The diffraction experiment provides intensities. Phases must be accessed indirectly by molecular replacement, or by experimental phasing. A popular method for crystallising membrane proteins employs a lipid cubic mesophase (the in meso method). Monoolein is the most popular lipid for in meso crystallisation. Invariably, the lipid co-crystallises with the protein recapitulating the biomembrane from whence it came. We reasoned that such a lipid bearing a heavy atom could be used for experimental phasing. In this study, we replaced half the monoolein in the mesophase with a seleno-labelled analogue (Se-MAG), which has a selenium atom in the fatty acyl chain of the lipid. The lipid mixture formed the cubic mesophase and grew crystals by the in meso method of the alginate transporter, AlgE, and the lipoprotein N-acyltransferase, Lnt. Se-MAGs co-crystallised with both proteins and were used to obtain phases for high-resolution structure determination by the selenium single-wavelength anomalous diffraction method. The use of such a mixed lipid system may prove to be a general strategy for the experimental phasing part of crystallographic structure determination of membrane proteins that crystallise via the in meso method.

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Section: Discussionmentioning

confidence: 99%

Se-MAG Is a Convenient Additive for Experimental Phasing and Structure Determination of Membrane Proteins Crystallised by the Lipid Cubic Phase (In Meso) Method

Boland,

Huang,

Shanker Kaki

et al. 2023

Crystals

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show abstract

“…In the review by Shinya Hanashima et al [ 12 ], molecular level structural analyses of the interaction between membrane proteins and lipids or detergents that constitute biological or artificial model membranes are shown to be important for understanding the functions and physicochemical properties of membrane proteins and biomembranes. A determination of membrane protein structures at the molecular level is much more difficult when compared with that of soluble proteins, but the new technical development has accelerated the elucidation of the structure–function relationship of membrane proteins.…”

mentioning

confidence: 99%

Advanced Research on Structure–Function Relationships of Membrane Proteins

Naito

Kawamura

2022

Membranes

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Rationalizing the Optimization of Detergents for Membrane Protein Purification

Urner

Junge

Fiorentino

et al. 2023

Chemistry A European J

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Membrane protein purification by means of detergents is key to isolating membrane‐bound therapeutic targets. The role of the detergent structure in this process, however, is not well understood. Detergents are optimized empirically, leading to failed preparations, and thereby raising costs. Here we evaluate the utility of the hydrophilic‐lipophilic balance (HLB) concept, which was introduced by Griffin in 1949, for guiding the optimization of the hydrophobic tail in first‐generation, dendritic oligoglycerol detergents ([G1] OGDs). Our findings deliver qualitative HLB guidelines for rationalizing the optimization of detergents. Moreover, [G1] OGDs exhibit strongly delipidating properties, regardless of the structure of the hydrophobic tail, which delivers a methodological enabling step for investigating binding strengths of endogenous lipids and their role for membrane protein oligomerization. Our findings will facilitate the analysis of challenging drug targets in the future.

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Heavy Atom Detergent/Lipid Combined X-ray Crystallography for Elucidating the Structure-Function Relationships of Membrane Proteins

Cited by 7 publications

References 76 publications

Se-MAG Is a Convenient Additive for Experimental Phasing and Structure Determination of Membrane Proteins Crystallised by the Lipid Cubic Phase (In Meso) Method

Se-MAG Is a Convenient Additive for Experimental Phasing and Structure Determination of Membrane Proteins Crystallised by the Lipid Cubic Phase (In Meso) Method

Advanced Research on Structure–Function Relationships of Membrane Proteins

Rationalizing the Optimization of Detergents for Membrane Protein Purification

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