Maltose neopentyl glycol-3 (MNG-3) analogues for membrane protein study

Cho, Kyung Ho; Husri, Mohd Naqiuddin; Amin, Anowarul; Gotfryd, Kamil; Lee, Ho Jin; Go, Juyeon; Kim, Jinwoong; Løland, Claus J.; Guan, Lan; Byrne, Bernadette; Chae, Pil Seok

doi:10.1039/c5an00240k

Cited by 44 publications

(53 citation statements)

References 68 publications

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“…Equal-volume solutions were analyzed by SDS-PAGE and immunoblotted with Penta-His-HRP antibody. The thermostability data of WT MelB St 27 and MelB Ec 1 in MNG-3 have been published.…”

Section: Methodsmentioning

confidence: 99%

Effect of Detergents on Galactoside Binding by Melibiose Permeases

et al. 2015

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The effect of various detergents on the stability and function of melibiose permeases of Escherichia coli (MelBEc) or Salmonella typhimurium (MelBSt) were studied. In n-dodecyl-β-d-maltoside (DDM) or n-undecyl-β-d-maltoside (UDM), WT MelBSt binds melibiose with an affinity similar to that in the membrane. However, with WT MelBEc or MelBSt mutants (Arg141→Cys, Arg295→Cys or Arg363→Cys), galactoside binding is not detected in these detergents, but binding to the phosphotransferase protein IIAGlc is maintained. In the amphiphiles lauryl maltose neopentyl glycol (MNG-3) or glyco-diosgenin (GDN), galactoside binding with all the MelB proteins is observed, with slightly reduced affinities. MelBSt is more thermostable than MelBEc, and the thermostability of either MelB is largely increased in MNG-3 or GDN. Therefore, the functional defect with DDM or UDM likely results from relative instability of the sensitive MelB proteins, and stability, as well as galactoside binding, is retained in MNG-3 or GDN. Furthermore, isothermal titration calorimetry of melibiose binding with MelBSt shows that the favorable entropic contribution to the binding free energy is decreased in MNG-3, indicating that the conformational dynamics of MelB is restricted in this detergent.

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“…Equal-volume solutions were analyzed by SDS-PAGE and immunoblotted with Penta-His-HRP antibody. The thermostability data of WT MelB St 27 and MelB Ec 1 in MNG-3 have been published.…”

Section: Methodsmentioning

confidence: 99%

Effect of Detergents on Galactoside Binding by Melibiose Permeases

et al. 2015

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“…Although conventional detergents such as DDM are capable of making membrane proteins soluble in aqueous solution, and much has been achieved using them, they often do not stabilise the membrane protein efficiently and can lead to protein aggregation [11]. To overcome these problems, novel detergents such as GNG, MNG and calixarenes have been produced that enhance stability [30,34,35]. Whilst the number of structures obtained using these novel detergents is still much lower than with conventional detergents, this may just be a matter of time.…”

Section: Discussionmentioning

confidence: 99%

“…Use of GNG-3 has help to produce the structure of Na + -pumping pyrophosphatase [32] and human aquaporin 2 (AQZ 2) [33]. MNG-3 has demonstrated great solubility and stability characteristics and its use facilitated the determination of the high resolution structure of more than 10 GPCRs [34]. Although it should be noted that for many of these structures, mutational/fusion protein approaches were also used in conjunction with the novel detergents.…”

Section: Glucose Neopentyl Glycol and Maltose Neopentyl Glycolmentioning

confidence: 99%

Overcoming bottlenecks in the membrane protein structural biology pipeline

Hardy

Bill

Jawhari³

et al. 2016

Biochemical Society Transactions

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Correspondence to: Alice Rothnie (+44 121 204 4013 a.rothnie@aston.ac.uk) or Anass Jawhari (+33 649 555 606 ajawhari@calixar.com) Key words:Membrane proteins Structural biology SMALP Calixarenes MNG Solubilisation Abbreviations:EM-Electron Microscopy GPCR-G protein-coupled receptors GNG-Glucose Neopentyl Glycol MSP -Membrane Scaffold Protein MNG-Maltose Neopentyl Glycol NMR-Nuclear magnetic resonance SMA-Styrene Maleic Acid SMALPs -SMA Lipid Particles AbstractMembrane proteins account for a third of the eukaryotic proteome, but are greatly underrepresented in the Protein Data Bank. Unfortunately, recent technological advances in X-ray crystallography and electron microscopy cannot account for the poor solubility and stability of membrane protein samples. A limitation of conventional detergent-based methods is that detergent molecules destabilize membrane proteins, leading to their aggregation. The use of orthologues, mutants and fusion tags has helped improve protein stability, but at the expense of not working with the sequence of interest. Novel detergents such as GNG, MNG and calixarene-based detergents can improve protein stability without compromising their solubilising properties. SMALPs focus on retaining the native lipid bilayer of a membrane protein during purification and biophysical analysis.Overcoming bottlenecks in the membrane protein structural biology pipeline, primarily by maintaining protein stability, will facilitate the elucidation of many more membrane protein structures in the near future.

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“…In the case of LeuT, a previous study has indicated that MNG-3 is slightly superior to DDM at maintaining transporter stability. [27] In contrast, some of the RGAs conferred substantially enhanced stability to the transporter under the same conditions. RGA-C11/C13 produced substantially smaller protein–detergent complex sizes compared to MNG-3, similar to that formed by DDM.…”

mentioning

confidence: 99%

Resorcinarene‐Based Facial Glycosides: Implication of Detergent Flexibility on Membrane‐Protein Stability

Hussain

Tikhonova

et al. 2017

Chemistry A European J

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As a membrane-mimetic system, detergent micelles are popularly used to extract membrane proteins from lipid environments and to maintain their solubility and stability in an aqueous medium. However, many membrane proteins encapsulated in conventional detergents tend to undergo structural degradation during extraction and purification, thus necessitating the development of new agents with enhanced properties. In the current study, two classes of new amphiphiles are introduced, resorcinarene-based glucoside and maltoside amphiphiles (designated RGAs and RMAs, respectively), for which the alkyl chains are facially segregated from the carbohydrate head groups. Of these facial amphiphiles, two RGAs (RGA-C11 and RGA-C13) conferred markedly enhanced stability to four tested membrane proteins compared to a gold-standard conventional detergent. The relatively high water solubility and micellar stability of the RGAs compared to the RMAs, along with their generally favourable behaviours for membrane protein stabilisation described here, are likely to be, at least in part, a result of the high conformational flexibility of these glucosides. This study suggests that flexibility could be an important factor in determining the suitability of new detergents for membrane protein studies.

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Maltose neopentyl glycol-3 (MNG-3) analogues for membrane protein study

Cited by 44 publications

References 68 publications

Effect of Detergents on Galactoside Binding by Melibiose Permeases

Effect of Detergents on Galactoside Binding by Melibiose Permeases

Overcoming bottlenecks in the membrane protein structural biology pipeline

Resorcinarene‐Based Facial Glycosides: Implication of Detergent Flexibility on Membrane‐Protein Stability

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