Controlled In Meso Phase Crystallization – A Method for the Structural Investigation of Membrane Proteins

Kubíček, Jan; Schlesinger, Ramona; Baeken, Christian; Büldt, Georg; Schäfer, Frank; Labahn, Jörg

doi:10.1371/journal.pone.0035458

Cited by 13 publications

(15 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…From 1996 to present this in-meso crystallization method has been greatly improved [199–203] and it has seen numerous successes in elucidating mechanisms of action of several microbial rhodopsins [204–208], in defining the crystal structure of the photosynthetic reaction centre from Rhodobacter sphaeroides [209], the high-resolution details of human G protein-coupled receptors bound to ligands [210], and of other free or liganded human proteins, such as the human A2A Adenosine Receptor Bound to an Antagonist [211], the human histamine H1 receptor complex with doxepin [212], the human M2 muscarinic acetylcholine receptor bound to an antagonist [213], etc. Recently, many efforts have been done to obtain an automatic high-throughput mode to perform cubic phase experiments [214–217]. The protein containing LCP is used in crystallization experiments with the advantage of obtaining crystals that are stabilized through extensive contacts in both the hydrophilic and hydrophobic protein regions [196].…”

Section: Membrane Proteinsmentioning

confidence: 99%

An Overview of Biological Macromolecule Crystallization

Krauss

Merlino

Vergara

et al. 2013

IJMS

127

View full text Add to dashboard Cite

The elucidation of the three dimensional structure of biological macromolecules has provided an important contribution to our current understanding of many basic mechanisms involved in life processes. This enormous impact largely results from the ability of X-ray crystallography to provide accurate structural details at atomic resolution that are a prerequisite for a deeper insight on the way in which bio-macromolecules interact with each other to build up supramolecular nano-machines capable of performing specialized biological functions. With the advent of high-energy synchrotron sources and the development of sophisticated software to solve X-ray and neutron crystal structures of large molecules, the crystallization step has become even more the bottleneck of a successful structure determination. This review introduces the general aspects of protein crystallization, summarizes conventional and innovative crystallization methods and focuses on the new strategies utilized to improve the success rate of experiments and increase crystal diffraction quality.

show abstract

Section: Membrane Proteinsmentioning

confidence: 99%

An Overview of Biological Macromolecule Crystallization

Krauss

Merlino

Vergara

et al. 2013

IJMS

127

View full text Add to dashboard Cite

show abstract

“…Membrane protein crystallography structures are resolved from hydrated protein crystals that contain about 30–70% water by volume or by weight in monoolein‐water media . Although most water molecules in crystals are dynamic and invisible, multiple well‐coordinated H 2 O molecules are repeatedly resolved and emerge as integral structural components of TM protein complexes.…”

Section: Overcoming the Multivalent Detergent‐phobia By Understandingmentioning

confidence: 99%

Detergents: Friends not foes for high‐performance membrane proteomics toward precision medicine

Zhang

2016

Proteomics

View full text Add to dashboard Cite

Precision medicine, particularly therapeutics, emphasizes the atomic-precise, dynamic, and systems visualization of human membrane proteins and their endogenous modifiers. For years, bottom-up proteomics has grappled with removing and avoiding detergents, yet faltered at the therapeutic-pivotal membrane proteins, which have been tackled by classical approaches and are known for decades refractory to single-phase aqueous or organic denaturants. Hydrophobicity and aggregation commonly challenge tissue and cell lysates, biofluids, and enriched samples. Frequently, expected membrane proteins and peptides are not identified by shotgun bottom-up proteomics, let alone robust quantitation. This review argues the cause of this proteomic crisis is not detergents per se, but the choice of detergents. Recently, inclusion of compatible detergents for membrane protein extraction and digestion has revealed stark improvements in both quantitative and structural proteomics. This review analyzes detergent properties behind recent proteomic advances, and proposes that rational use of detergents may reconcile outstanding membrane proteomics dilemmas, enabling ultradeep coverage and minimal artifacts for robust protein and endogenous PTM measurements. The simplicity of detergent tools confers bottom-up membrane proteomics the sophistication toward precision medicine.

show abstract

“…At present, the in cubo method is widely used and applications for soluble proteins are expected. Recently, the method was extended to other mesophases in the lipid (monoolein)/water diagrams and led to a user‐friendly fast screening technology .…”

Section: Crystallogenesis In the Era Of Technologies And Structural Gmentioning

confidence: 99%

A historical perspective on protein crystallization from 1840 to the present day

Giegé

2013

FEBS J

110

View full text Add to dashboard Cite

Protein crystallization has been known since 1840 and can prove to be straightforward but, in most cases, it constitutes a real bottleneck. This stimulated the birth of the biocrystallogenesis field with both ‘practical’ and ‘basic’ science aims. In the early years of biochemistry, crystallization was a tool for the preparation of biological substances. Today, biocrystallogenesis aims to provide efficient methods for crystal fabrication and a means to optimize crystal quality for X‐ray crystallography. The historical development of crystallization methods for structural biology occurred first in conjunction with that of biochemical and genetic methods for macromolecule production, then with the development of structure determination methodologies and, recently, with routine access to synchrotron X‐ray sources. Previously, the identification of conditions that sustain crystal growth occurred mostly empirically but, in recent decades, this has moved progressively towards more rationality as a result of a deeper understanding of the physical chemistry of protein crystal growth and the use of idea‐driven screening and high‐throughput procedures. Protein and nucleic acid engineering procedures to facilitate crystallization, as well as crystallization methods in gelled‐media or by counter‐diffusion, represent recent important achievements, although the underlying concepts are old. The new nanotechnologies have brought a significant improvement in the practice of protein crystallization. Today, the increasing number of crystal structures deposited in the Protein Data Bank could mean that crystallization is no longer a bottleneck. This is not the case, however, because structural biology projects always become more challenging and thereby require adapted methods to enable the growth of the appropriate crystals, notably macromolecular assemblages.

show abstract

Controlled In Meso Phase Crystallization – A Method for the Structural Investigation of Membrane Proteins

Cited by 13 publications

References 34 publications

An Overview of Biological Macromolecule Crystallization

An Overview of Biological Macromolecule Crystallization

Detergents: Friends not foes for high‐performance membrane proteomics toward precision medicine

A historical perspective on protein crystallization from 1840 to the present day

Contact Info

Product

Resources

About