Peculiarities of thermal denaturation of OmpF porin from Yersinia ruckeri

2017 ◽  
Vol 13 (9) ◽  
pp. 1854-1862 ◽  
Author(s):  
Olga D. Novikova ◽  
Dmitry K. Chistyulin ◽  
Valentina A. Khomenko ◽  
Evgeny V. Sidorin ◽  
Natalya Yu. Kim ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Thermal Denaturation ◽  
Water Soluble ◽  
Yersinia Ruckeri ◽  
Multidomain Proteins ◽  
Multistage Process ◽  
Ompf Porin ◽  
Irreversible Denaturation ◽  
Detergent Solutions

Irreversible denaturation of membrane proteins in detergent solutions is similar to unfolding of water-soluble multidomain proteins and represents a complex, multistage process.

Block poly(Ala)-poly(Lys). A water-soluble model for intrinsic membrane proteins?

1979 ◽  
Vol 557 (2) ◽  
pp. 331-339 ◽  
Author(s):  
L. Vitello ◽  
G.C. Kresheck ◽  
R.J. Albers ◽  
J.E. Erman ◽  
G. Vanderkooi
Keyword(s):  
Membrane Proteins ◽  
Water Soluble ◽  
Soluble Model

scholarly journals Membrane Proteins Are Dramatically Less Conserved than Water-Soluble Proteins across the Tree of Life

2016 ◽  
Vol 33 (11) ◽  
pp. 2874-2884 ◽  
Author(s):  
Victor Sojo ◽  
Christophe Dessimoz ◽  
Andrew Pomiankowski ◽  
Nick Lane
Keyword(s):  
Membrane Proteins ◽  
Soluble Proteins ◽  
Tree Of Life ◽  
Water Soluble

scholarly journals Membrane proteins: always an insoluble problem?

2016 ◽  
Vol 44 (3) ◽  
pp. 790-795 ◽  
Author(s):  
Andrea E. Rawlings
Keyword(s):  
Membrane Proteins ◽  
Lipid Membranes ◽  
Drug Targets ◽  
Functional Characterization ◽  
Water Soluble ◽  
High Yield ◽  
Cellular Processes ◽  
Native Sequence ◽  
Protein Research ◽  
New Methodologies

Membrane proteins play crucial roles in cellular processes and are often important pharmacological drug targets. The hydrophobic properties of these proteins make full structural and functional characterization challenging because of the need to use detergents or other solubilizing agents when extracting them from their native lipid membranes. To aid membrane protein research, new methodologies are required to allow these proteins to be expressed and purified cheaply, easily, in high yield and to provide water soluble proteins for subsequent study. This mini review focuses on the relatively new area of water soluble membrane proteins and in particular two innovative approaches: the redesign of membrane proteins to yield water soluble variants and how adding solubilizing fusion proteins can help to overcome these challenges. This review also looks at naturally occurring membrane proteins, which are able to exist as stable, functional, water soluble assemblies with no alteration to their native sequence.

Elucidation of cosolvent effects thermostabilizing water-soluble and membrane proteins

2020 ◽  
Vol 301 ◽  
pp. 112403 ◽  
Author(s):  
Satoshi Yasuda ◽  
Kazuki Kazama ◽  
Tomoki Akiyama ◽  
Masahiro Kinoshita ◽  
Takeshi Murata
Keyword(s):  
Membrane Proteins ◽  
Water Soluble ◽  
Cosolvent Effects

Notes: Thermal Lability of Membrane Proteins of Age Separated Erythrocytes as Studied by Electron Spin Resonance Spin Label Technique

1987 ◽  
Vol 42 (11-12) ◽  
pp. 1343-1344 ◽  
Author(s):  
Grzegorz Bartosz ◽  
Gabriele Christ ◽  
Harald Bosse ◽  
Roland Stephan ◽  
Helmut Gärtner
Keyword(s):  
Electron Spin Resonance ◽  
Membrane Proteins ◽  
Erythrocyte Membrane ◽  
Electron Spin ◽  
Spin Label ◽  
Thermal Denaturation ◽  
Spin Resonance ◽  
Resonance Spin ◽  
First Time ◽  
Erythrocyte Membrane Proteins

Thermal lability of bovine erythrocyte membrane proteins was studied by electron spin resonance using maleimide spin label. The temperature of the sample during measurements could be varied for the first time be­ tween 0 and 60 °C with an accuracy of ± 0.1 °C. Our results show that “old” erythrocyte membrane proteins are less stable against thermal denaturation then “young” cells.

scholarly journals Effects of ionizing radiations on proteins. Evidence of non-random fragmentations and a caution in the use of the method for determination of molecular mass

1990 ◽  
Vol 267 (2) ◽  
pp. 431-439 ◽  
Author(s):  
M Le Maire ◽  
L Thauvette ◽  
B de Foresta ◽  
A Viel ◽  
G Beauregard ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Molecular Mass ◽  
Three Dimensional ◽  
Water Soluble ◽  
Dimensional Structure ◽  
Intact Protein ◽  
Aspartate Transcarbamylase ◽  
Ionizing Radiations ◽  
Polypeptide Chains ◽  
Best Fit

We have reinvestigated the use of ionizing radiations to measure the molecular mass of water-soluble or membrane proteins. The test was performed by using the most straightforward aspect of the technique, which consists of SDS/PAGE analysis of the protein-fragmentation process. We found that exposure of purified standard proteins to increasing doses of ionizing radiation causes progressive fragmentation of the native protein into defined peptide patterns. The coloured band corresponding to the intact protein was measured on the SDS gel as a function of dose to determine the dose (D37.t) corresponding to 37% of the initial amount of unfragmented protein deposited on the gel. This led to a calibration curve between 1/D37.t and the known molecular mass of the standard proteins whose best fit gave Mr = 1.77 x 10(6)/D37.t at -78 degrees C, i.e. 35% higher than the generally accepted value at that temperature obtained from inactivation studies. However, we have to conclude that this method is useless to determine the state of aggregation of a protein, since, for all the oligomers tested, the best fit was obtained by using the protomeric molecular mass, suggesting that there is no energy transfer between promoters. Furthermore, SDS greatly increases the fragmentation rate of proteins, which suggests additional calibration problems for membrane proteins in detergent or in the lipid bilayer. But the main drawback of the technique arises from our observation that some proteins behaved anomalously, leading to very large errors in the apparent target size as compared with true molecular mass (up to 100%). It is thus unreliable to apply the radiation method for absolute molecular-mass determination. We then focused on the novel finding that discrete fragmentation of proteins occurs at preferential sites, and this was studied in more detail with aspartate transcarbamylase. N-Terminal sequencing of several radiolysis fragments of the catalytic chain of the enzyme revealed that breaks along the polypeptide chains are localized close to the C-terminal end. Examination of the three-dimensional structure of aspartate transcarbamylase suggests that radiolysis sites (fragile bonds) might be localized in connecting loops.

scholarly journals Making water-soluble integral membrane proteins in vivo using an amphipathic protein fusion strategy

2015 ◽  
Vol 6 (1) ◽  
Author(s):  
Dario Mizrachi ◽  
Yujie Chen ◽  
Jiayan Liu ◽  
Hwei-Ming Peng ◽  
Ailong Ke ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Water Soluble ◽  
Integral Membrane Proteins ◽  
Protein Fusion ◽  
Fusion Strategy

scholarly journals The Peptidisc, a simple method for stabilizing membrane proteins in detergent-free solution

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Michael Luke Carlson ◽  
John William Young ◽  
Zhiyu Zhao ◽  
Lucien Fabre ◽  
Daniel Jun ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Cost Effective ◽  
Water Soluble ◽  
Protein Assemblies ◽  
Simple Method ◽  
Helical Peptide ◽  
Biological Studies ◽  
Multiple Copies ◽  
Exposed Part

Membrane proteins are difficult to work with due to their insolubility in aqueous solution and quite often their poor stability in detergent micelles. Here, we present the peptidisc for their facile capture into water-soluble particles. Unlike the nanodisc, which requires scaffold proteins of different lengths and precise amounts of matching lipids, reconstitution of detergent solubilized proteins in peptidisc only requires a short amphipathic bi-helical peptide (NSPr) and no extra lipids. Multiple copies of the peptide wrap around to shield the membrane-exposed part of the target protein. We demonstrate the effectiveness of this ‘one size fits all’ method using five different membrane protein assemblies (MalFGK2, FhuA, SecYEG, OmpF, BRC) during ‘on-column’, ‘in-gel’, and ‘on-bead’ reconstitution embedded within the membrane protein purification protocol. The peptidisc method is rapid and cost-effective, and it may emerge as a universal tool for high-throughput stabilization of membrane proteins to advance modern biological studies.

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