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

Stetsenko, Artem; Guskov, Albert

doi:10.3390/cryst7070197

Cited by 111 publications

(68 citation statements)

References 88 publications

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“…Considering the findings from the solubilization screen and F-SEC analysis, as well as previous reports demonstrating that fos-cholines can be detrimental to membrane proteins [55,56], and the fact that both DDM and DM are popular detergents for structural and functional studies (also of TRP channels) [9,57], we decided to use primarily these milder surfactants for the subsequent large-scale purifications.…”

Section: Resultsmentioning

confidence: 99%

Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

Zhang

Wang

Klærke

et al. 2019

Cells

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(1) Background: Human transient receptor potential (TRP) channels constitute a large family of ion-conducting membrane proteins that allow the sensation of environmental cues. As the dysfunction of TRP channels contributes to the pathogenesis of many widespread diseases, including cardiac disorders, these proteins also represent important pharmacological targets. TRP channels are typically produced using expensive and laborious mammalian or insect cell-based systems. (2) Methods: We demonstrate an alternative platform exploiting the yeast Saccharomyces cerevisiae capable of delivering high yields of functional human TRP channels. We produce 11 full-length human TRP members originating from four different subfamilies, purify a selected subset of these to a high homogeneity and confirm retained functionality using TRPM8 as a model target. (3) Results: Our findings demonstrate the potential of the described production system for future functional, structural and pharmacological studies of human TRP channels.

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Section: Resultsmentioning

confidence: 99%

Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

Zhang

Wang

Klærke

et al. 2019

Cells

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“…MP purification is traditionally focused on identification of the most suitable detergent for purification before exchange into a shorter chain detergent prior to crystallisation. Detergents are still among the most common tools used to extract MPs from membranes, and choice of detergent is a critical step when trying to purify an MP for crystallisation or cryo-EM [93,94]. Over the last twenty years, new generations of detergents have been developed.…”

Section: Use Of Encapsulation Agents To Stabilise Membrane Proteinsmentioning

confidence: 99%

“…Consequently, type II crystals have a lower solvent content and are, therefore, more robust [171]. Selection of the right detergents for direct crystallisation is necessary to obtain well diffracting crystals [94,102,185]. Detergents with shorter chains form smaller micelles that result in tighter crystal lattice packing and improved diffraction compared to detergents with longer acyl chains, although these tend to be more efficient for solubilisation and stabilisation [102,186,187].…”

Section: Membrane Protein Crystallisation and Data Collectionmentioning

confidence: 99%

Changes in Membrane Protein Structural Biology

Birch

Cheruvara

Gamage

et al. 2020

Biology

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Membrane proteins are essential components of many biochemical processes and are important pharmaceutical targets. Membrane protein structural biology provides the molecular rationale for these biochemical process as well as being a highly useful tool for drug discovery. Unfortunately, membrane protein structural biology is a difficult area of study due to low protein yields and high levels of instability especially when membrane proteins are removed from their native environments. Despite this instability, membrane protein structural biology has made great leaps over the last fifteen years. Today, the landscape is almost unrecognisable. The numbers of available atomic resolution structures have increased 10-fold though advances in crystallography and more recently by cryo-electron microscopy. These advances in structural biology were achieved through the efforts of many researchers around the world as well as initiatives such as the Membrane Protein Laboratory (MPL) at Diamond Light Source. The MPL has helped, provided access to and contributed to advances in protein production, sample preparation and data collection. Together, these advances have enabled higher resolution structures, from less material, at a greater rate, from a more diverse range of membrane protein targets. Despite this success, significant challenges remain. Here, we review the progress made and highlight current and future challenges that will be overcome.

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“…To address these shortcomings, new techniques have been developed to test the mechanical properties of the scaffold (Den and to measure the amount of decellularization reagents remaining after decellularization (Dettin et al, 2017). Knowledge of the critical micelle concentration of detergents (Stetsenko & Guskov, 2017) should aid in the development of standardized decellularization solutions that prevent micelle formation and thereby the risk of deleterious protein extraction. A recent study has established a threshold for the maximal amount of residual antigenicity of an unfixed xenogeneic scaffold to overcome the adaptive humoral immune response after transplantation.…”

Section: Standardization Of Decellularizationmentioning

confidence: 99%

Limitations of recellularized biological scaffolds for human transplantation

Bilodeau

Goltsis

Rogers

et al. 2020

J Tissue Eng Regen Med

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A shortage of donor organs for transplantation and the dependence of the recipients on immunosuppressive therapy have motivated researchers to consider alternative regenerative approaches. The answer may reside in acellular scaffolds generated from cadaveric human and animal tissues. Acellular scaffolds are expected to preserve the architectural and mechanical properties of the original organ, permitting cell attachment, growth, and differentiation. Although theoretically, the use of acellular scaffolds for transplantation should pose no threat to the recipient's immune system, experimental data have revealed significant immune responses to allogeneic and xenogeneic transplanted scaffolds. Herein, we review the various factors of the scaffold that could trigger an inflammatory and/or immune response, thereby compromising its use for human transplant therapy. In addition, we provide an overview of the major cell types that have been considered for recellularization of the scaffold and their potential contribution to triggering an immune response.

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

Cited by 111 publications

References 88 publications

Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

Purification of Functional Human TRP Channels Recombinantly Produced in Yeast

Changes in Membrane Protein Structural Biology

Limitations of recellularized biological scaffolds for human transplantation

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