Tailored nanodisc immobilization for one‐step purification and reconstitution of cytochrome P450: A tool for membrane proteins’ hard cases

2021 ◽  
Author(s):  
Lan Zhao ◽  
Jiaoli Tao ◽  
Yongdong Huang ◽  
Kai Zhu ◽  
Yuxiang Du ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Membrane Proteins ◽  
One Step ◽  
Hard Cases

scholarly journals DirectMX – One-Step Reconstitution of Membrane Proteins From Crude Cell Membranes Into Salipro Nanoparticles

2020 ◽  
Vol 8 ◽  
Author(s):  
Pilar Lloris-Garcerá ◽  
Stefan Klinter ◽  
Liuhong Chen ◽  
Michael J. Skynner ◽  
Robin Löving ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Cell Membranes ◽  
One Step

Comparison of cytochrome P450 2A6 polymorphism frequencies in Caucasians and African-Americans using a new one-step PCR-RFLP genotyping method

Toxicology ◽  
2001 ◽  
Vol 168 (3) ◽  
pp. 259-268 ◽  
Author(s):  
Thilo Paschke ◽  
Michelle Riefler ◽  
Annette Schuler-Metz ◽  
Lucie Wolz ◽  
Gerhard Scherer ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
African Americans ◽  
Genotyping Method ◽  
Pcr Rflp ◽  
Cytochrome P450 2A6 ◽  
One Step

scholarly journals Retention of membrane proteins by the endoplasmic reticulum.

1985 ◽  
Vol 101 (5) ◽  
pp. 1724-1732 ◽  
Author(s):  
R Brands ◽  
M D Snider ◽  
Y Hino ◽  
S S Park ◽  
H V Gelboin ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Endoplasmic Reticulum ◽  
Cytochrome P450 ◽  
Membrane Proteins ◽  
Nuclear Envelope ◽  
Smooth Endoplasmic Reticulum ◽  
Membrane Glycoprotein ◽  
Glucosidase Ii ◽  
Er Membrane

We have used a monoclonal antibody specific for a hydrocarbon-induced cytochrome P450 to localize, by electron microscopy, the epitope-specific cytochrome P450. The cytochrome was found in the rough and smooth endoplasmic reticulum (ER) and the nuclear envelope of hepatocytes. Significant quantities of cytochrome P450 were not found in Golgi stacks. We also could not find any evidence of Golgi-associated processing of the Asn-linked oligosaccharide chains of two well-characterized ER membrane glycoprotein enzymes (glucosidase II and hexose-6-phosphate dehydrogenase), or of the oligosaccharides attached to the bulk of the glycoproteins of the ER membrane. We conclude that these ER membrane proteins are efficiently retained during a process of highly selective export from this organelle.

AFM Study of Membrane Proteins, Cytochrome P450 2B4, and NADPH–Cytochrome P450 Reductase and Their Complex Formation

1999 ◽  
Vol 371 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Olga I. Kiselyova ◽  
Igor V. Yaminsky ◽  
Yuri D. Ivanov ◽  
Irina P. Kanaeva ◽  
Vadim Yu. Kuznetsov ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Membrane Proteins ◽  
Complex Formation ◽  
Cytochrome P450 Reductase ◽  
Nadph Cytochrome ◽  
P450 Reductase ◽  
Cytochrome P450 2B4 ◽  
Nadph Cytochrome P450 Reductase

scholarly journals One-step isolation of plasma membrane proteins using magnetic beads with immobilized concanavalin A

2008 ◽  
Vol 62 (2) ◽  
pp. 223-229 ◽  
Author(s):  
Yu-Chen Lee ◽  
Gregory Block ◽  
Huiwen Chen ◽  
Emma Folch-Puy ◽  
Robert Foronjy ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Concanavalin A ◽  
Magnetic Beads ◽  
Plasma Membrane Proteins ◽  
One Step

scholarly journals A one-step purification of membrane proteins using a high efficiency immunomatrix.

1982 ◽  
Vol 257 (18) ◽  
pp. 10766-10769 ◽  
Author(s):  
C Schneider ◽  
R A Newman ◽  
D R Sutherland ◽  
U Asser ◽  
M F Greaves
Keyword(s):  
Membrane Proteins ◽  
High Efficiency ◽  
One Step

scholarly journals Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis

2021 ◽  
Author(s):  
Anna Higgins ◽  
Alex Flynn ◽  
Anaïs Marconnet ◽  
Laura Musgrove ◽  
Vincent Postis ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Membrane Proteins ◽  
Amphiphilic Polymers ◽  
Cellular Growth ◽  
Functional Study ◽  
Direct Extraction ◽  
Detergent Extraction ◽  
High Resolution Structure ◽  
One Step ◽  
Resolution Structure

Abstract Membrane proteins are essential for cellular growth and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them from the membrane creates significant challenges in their structural and functional study. Although amphipols have been very effective for single-particle electron cryo-microscopy (cryoEM) and mass spectrometry, they rely on initial detergent extraction before exchange into the amphipol environment. Therefore, circumventing this pre-requirement would be a significant advantage. Here we use a novel type of amphipol: a cycloalkane-modified amphiphile polymer (CyclAPol) to extract Escherichia coli AcrB directly from the membrane and demonstrate that the protein can be isolated in a one-step purification with the resultant cryoEM structure achieving 3.2 Å resolution. Together this work shows that cycloalkane amphipols provide a powerful detergent-free approach for the study of membrane proteins allowing native extraction and high-resolution structure determination by cryoEM.

scholarly journals One-step construction of circularized nanodiscs using SpyCatcher-SpyTag

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Shanwen Zhang ◽  
Qian Ren ◽  
Scott J. Novick ◽  
Timothy S. Strutzenberg ◽  
Patrick R. Griffin ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Synaptic Transmission ◽  
Lipid Bilayers ◽  
Structural Biology ◽  
Viral Entry ◽  
Vesicle Fusion ◽  
Structural Studies ◽  
Lipid Interactions ◽  
Complex Membrane ◽  
One Step

AbstractCircularized nandiscs (cNDs) exhibit superb monodispersity and have the potential to transform functional and structural studies of membrane proteins. In particular, cNDs can stabilize large patches of lipid bilayers for the reconstitution of complex membrane biochemical reactions, enabling the capture of crucial intermediates involved in synaptic transmission and viral entry. However, previous methods for building cNDs require multiple steps and suffer from low yields. We herein introduce a simple, one-step approach to ease the construction of cNDs using the SpyCatcher-SpyTag technology. This approach increases the yield of cNDs by over 10-fold and is able to rapidly generates cNDs with diameters ranging from 11 to over 100 nm. We demonstrate the utility of these cNDs for mechanistic interrogations of vesicle fusion and protein-lipid interactions that are unattainable using small nanodiscs. Together, the remarkable performance of SpyCatcher-SpyTag in nanodisc circularization paves the way for the use of cNDs in membrane biochemistry and structural biology.

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