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Proceedings of the National Academy of... May 2023Integral membrane protein structure determination traditionally requires extraction from cell membranes using detergents or polymers. Here, we describe the isolation and...
Integral membrane protein structure determination traditionally requires extraction from cell membranes using detergents or polymers. Here, we describe the isolation and structure determination of proteins in membrane vesicles derived directly from cells. Structures of the ion channel Slo1 from total cell membranes and from cell plasma membranes were determined at 3.8 Å and 2.7 Å resolution, respectively. The plasma membrane environment stabilizes Slo1, revealing an alteration of global helical packing, polar lipid, and cholesterol interactions that stabilize previously unresolved regions of the channel and an additional ion binding site in the Ca regulatory domain. The two methods presented enable structural analysis of both internal and plasma membrane proteins without disrupting weakly interacting proteins, lipids, and cofactors that are essential to biological function.
Topics: Membrane Proteins; Cell Membrane; Ion Channels; Binding Sites
PubMed: 37098056
DOI: 10.1073/pnas.2302325120 -
Journal of the Royal Society, Interface Apr 2018This review highlights recent development of biosensors that use the functions of membrane proteins. Membrane proteins are essential components of biological membranes... (Review)
Review
This review highlights recent development of biosensors that use the functions of membrane proteins. Membrane proteins are essential components of biological membranes and have a central role in detection of various environmental stimuli such as olfaction and gustation. A number of studies have attempted for development of biosensors using the sensing property of these membrane proteins. Their specificity to target molecules is particularly attractive as it is significantly superior to that of traditional human-made sensors. In this review, we classified the membrane protein-based biosensors into two platforms: the lipid bilayer-based platform and the cell-based platform. On lipid bilayer platforms, the membrane proteins are embedded in a lipid bilayer that bridges between the protein and a sensor device. On cell-based platforms, the membrane proteins are expressed in a cultured cell, which is then integrated in a sensor device. For both platforms we introduce the fundamental information and the recent progress in the development of the biosensors, and remark on the outlook for practical biosensing applications.
Topics: Animals; Biosensing Techniques; Cell Membrane; HEK293 Cells; Humans; Lipid Bilayers; Membrane Proteins; MicroRNAs; Models, Molecular; Protein Engineering; Saccharomyces cerevisiae; Signal Transduction; Smell; Spodoptera; Xenopus laevis
PubMed: 29669891
DOI: 10.1098/rsif.2017.0952 -
The Journal of Biological Chemistry Mar 2018My scientific career has taken me from chemistry, via theoretical physics and bioinformatics, to molecular biology and even structural biology. Along the way,...
My scientific career has taken me from chemistry, via theoretical physics and bioinformatics, to molecular biology and even structural biology. Along the way, serendipity led me to work on problems such as the identification of signal peptides that direct protein trafficking, membrane protein biogenesis, and cotranslational protein folding. I've had some great collaborations that came about because of a stray conversation or from following up on an interesting paper. And I've had the good fortune to be asked to sit on the Nobel Committee for Chemistry, where I am constantly reminded of the amazing pace and often intricate history of scientific discovery. Could I have planned this? No way! I just went with the flow ….
Topics: Animals; Chemical Engineering; Computational Biology; History, 20th Century; History, 21st Century; Humans; Lipid Bilayers; Membrane Proteins; Models, Molecular; Molecular Biology; New York City; Physics; Protein Folding; Protein Sorting Signals; Signal Transduction; Sweden
PubMed: 29523692
DOI: 10.1074/jbc.X118.001958 -
Biochimica Et Biophysica Acta Jan 2014
Topics: Biophysics; Ligands; Membrane Proteins; Protein Binding; Protein Conformation
PubMed: 24206896
DOI: 10.1016/j.bbamem.2013.09.013 -
Biochemical Society Transactions Oct 2019The construction of artificial membrane proteins from first principles is of fundamental interest and holds considerable promise for new biotechnologies. This review... (Review)
Review
The construction of artificial membrane proteins from first principles is of fundamental interest and holds considerable promise for new biotechnologies. This review considers the potential advantages of adopting a strictly minimalist approach to the process of membrane protein design. As well as the practical benefits of miniaturisation and simplicity for understanding sequence-structure-function relationships, minimalism should also support the abstract conceptualisation of membrane proteins as modular components for synthetic biology. These ideas are illustrated with selected examples that focus upon α-helical membrane proteins, and which demonstrate how such minimalist membrane proteins might be integrated into living biosystems.
Topics: Biotechnology; Membrane Proteins; Structure-Activity Relationship; Synthetic Biology
PubMed: 31671181
DOI: 10.1042/BST20190170 -
Proceedings of the National Academy of... Sep 2021Cell surface receptors are critical for cell signaling and constitute a quarter of all human genes. Despite their importance and abundance, receptor interaction networks...
Cell surface receptors are critical for cell signaling and constitute a quarter of all human genes. Despite their importance and abundance, receptor interaction networks remain understudied because of difficulties associated with maintaining membrane proteins in their native conformation and their typically weak interactions. To overcome these challenges, we developed an extracellular vesicle-based method for membrane protein display that enables purification-free and high-throughput detection of receptor-ligand interactions in membranes. We demonstrate that this platform is broadly applicable to a variety of membrane proteins, enabling enhanced detection of extracellular interactions over a wide range of binding affinities. We were able to recapitulate and expand the interactome for prominent members of the B7 family of immunoregulatory proteins such as PD-L1/CD274 and B7-H3/CD276. Moreover, when applied to the orphan cancer-associated fibroblast protein, LRRC15, we identified a membrane-dependent interaction with the tumor stroma marker TEM1/CD248. Furthermore, this platform enabled profiling of cellular receptors for target-expressing as well as endogenous extracellular vesicles. Overall, this study presents a sensitive and easy to use screening platform that bypasses membrane protein purification and enables characterization of interactomes for any cell surface-expressed target of interest in its native state.
Topics: Antigens, CD; Antigens, Neoplasm; B7 Antigens; B7-H1 Antigen; HEK293 Cells; Humans; Membrane Proteins; Protein Interaction Domains and Motifs
PubMed: 34531301
DOI: 10.1073/pnas.2025451118 -
Annual Review of Biomedical Engineering Jul 2016The majority of therapeutics target membrane proteins, accessible on the surface of cells, to alter cellular signaling. Cells use membrane proteins to transduce signals... (Review)
Review
The majority of therapeutics target membrane proteins, accessible on the surface of cells, to alter cellular signaling. Cells use membrane proteins to transduce signals into cells, transport ions and molecules, bind cells to a surface or substrate, and catalyze reactions. Newly devised technologies allow us to drug conventionally "undruggable" regions of membrane proteins, enabling modulation of protein-protein, protein-lipid, and protein-nucleic acid interactions. In this review, we survey the state of the art of high-throughput screening and rational design in drug discovery, and we evaluate the advances in biological understanding and technological capacity that will drive pharmacotherapy forward against unorthodox membrane protein targets.
Topics: Animals; Drug Discovery; High-Throughput Screening Assays; Humans; Membrane Proteins; Molecular Targeted Therapy; Protein Interaction Mapping
PubMed: 26863923
DOI: 10.1146/annurev-bioeng-092115-025322 -
The Journal of Biological Chemistry Jun 2015The YidC/Alb3/Oxa1 family functions in the insertion and folding of proteins in the bacterial cytoplasmic membrane, the chloroplast thylakoid membrane, and the... (Review)
Review
The YidC/Alb3/Oxa1 family functions in the insertion and folding of proteins in the bacterial cytoplasmic membrane, the chloroplast thylakoid membrane, and the mitochondrial inner membrane. All members share a conserved region composed of five transmembrane regions. These proteins mediate membrane insertion of an assorted group of proteins, ranging from respiratory subunits in the mitochondria and light-harvesting chlorophyll-binding proteins in chloroplasts to ATP synthase subunits in bacteria. This review discusses the YidC/Alb3/Oxa1 protein family as well as their function in membrane insertion and two new structures of the bacterial YidC, which suggest a mechanism for membrane insertion by this family of insertases.
Topics: Membrane Proteins; Substrate Specificity; beta-Fructofuranosidase
PubMed: 25947384
DOI: 10.1074/jbc.R115.638171 -
Biological Chemistry Jun 2019Membrane proteins are key players in the cell. Due to their hydrophobic nature they require solubilising agents such as detergents or membrane mimetics during... (Review)
Review
Membrane proteins are key players in the cell. Due to their hydrophobic nature they require solubilising agents such as detergents or membrane mimetics during purification and, consequently, are challenging targets in structural biology. In addition, their natural lipid environment is crucial for their structure and function further hampering their analysis. Alternative approaches are therefore required when the analysis by conventional techniques proves difficult. In this review, we highlight the broad application of mass spectrometry (MS) for the characterisation of membrane proteins and their interactions with lipids. We show that MS unambiguously identifies the protein and lipid components of membrane protein complexes, unravels their three-dimensional arrangements and further provides clues of protein-lipid interactions.
Topics: Binding Sites; Hydrophobic and Hydrophilic Interactions; Mass Spectrometry; Membrane Lipids; Membrane Proteins; Protein Binding; Protein Conformation; Solubility
PubMed: 30956223
DOI: 10.1515/hsz-2018-0443 -
Annual Review of Biochemistry Jun 2021Microorganisms contend with numerous and unusual chemical threats and have evolved a catalog of resistance mechanisms in response. One particularly ancient, pernicious... (Review)
Review
Microorganisms contend with numerous and unusual chemical threats and have evolved a catalog of resistance mechanisms in response. One particularly ancient, pernicious threat is posed by fluoride ion (F), a common xenobiotic in natural environments that causes broad-spectrum harm to metabolic pathways. This review focuses on advances in the last ten years toward understanding the microbial response to cytoplasmic accumulation of F, with a special emphasis on the structure and mechanisms of the proteins that microbes use to export fluoride: the CLC family of F/H antiporters and the Fluc/FEX family of F channels.
Topics: Antiporters; Chloride Channels; Cytoplasm; Fluorides; Ion Channels; Ion Transport; Membrane Proteins; Protein Conformation; Saccharomyces cerevisiae Proteins
PubMed: 33492991
DOI: 10.1146/annurev-biochem-071520-112507