Use of Chemical Chaperones in the Yeast Saccharomyces cerevisiae to Enhance Heterologous Membrane Protein Expression: High-Yield Expression and Purification of Human P-Glycoprotein

2000 ◽  
Vol 376 (1) ◽  
pp. 34-46 ◽  
Author(s):  
Robert A. Figler ◽  
Hiroshi Omote ◽  
Robert K. Nakamoto ◽  
Marwan K. Al-Shawi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Protein ◽  
Protein Expression ◽  
High Yield ◽  
Expression And Purification ◽  
Yeast Saccharomyces Cerevisiae ◽  
Chemical Chaperones ◽  
Membrane Protein Expression ◽  
P Glycoprotein

Database Study on the Expression and Purification of Membrane Proteins

2021 ◽  
Vol 28 ◽  
Author(s):  
Chen-Yan china Zhang ◽  
Shi-Qi Zhao ◽  
Shi-Long Zhang ◽  
Li-Heng Luo ◽  
Ding-Chang Liu ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Protein Expression ◽  
Drug Targets ◽  
Expression System ◽  
Data Bank ◽  
Hek293 Cells ◽  
Expression And Purification ◽  
Beta Barrel ◽  
Membrane Protein Expression

: Membrane proteins are crucial for biological processes, and many of them are important to drug targets. Understanding the three-dimensional structures of membrane proteins are essential to evaluate their bio function and drug design. High-purity membrane proteins are important for structural determination. Membrane proteins have low yields and are difficult to purify because they tend to aggregate. We summarized membrane protein expression systems, vectors, tags, and detergents, which have deposited in the Protein Data Bank (PDB) in recent four-and-a-half years. Escherichia coli is the most expression system for membrane proteins, and HEK293 cells are the most commonly cell lines for human membrane protein expression. The most frequently vectors are pFastBac1 for alpha-helical membrane proteins, pET28a for beta-barrel membrane proteins, and pTRC99a for monotopic membrane proteins. The most used tag for membrane proteins is the 6×His-tag. FLAG commonly used for alpha-helical membrane proteins, Strep and GST for beta-barrel and monotopic membrane proteins, respectively. The detergents and their concentrations used for alpha-helical, beta-barrel, and monotopic membrane proteins are different, and DDM is commonly used for membrane protein purification. It can guide the expression and purification of membrane proteins, thus contributing to their structure and bio function studying.

scholarly journals High-yield membrane protein expression from E. coli using an engineered outer membrane protein F fusion

2013 ◽  
Vol 22 (4) ◽  
pp. 434-443 ◽  
Author(s):  
Pin-Chuan Su ◽  
William Si ◽  
Deidre L. Baker ◽  
Bryan W. Berger
Keyword(s):  
Membrane Protein ◽  
Protein Expression ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
High Yield ◽  
E Coli ◽  
Membrane Protein Expression ◽  
Protein F ◽  
Outer Membrane Protein F

Membrane Protein Expression in Saccharomyces cerevisiae

2011 ◽  
pp. 37-73
Author(s):  
Zachary Britton ◽  
Carissa Young ◽  
Özge Can ◽  
Patrick McNeely ◽  
Andrea Naranjo ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Protein ◽  
Protein Expression ◽  
Membrane Protein Expression

scholarly journals Beverage–Drug Interaction: Effects of Green Tea Beverage Consumption on Atorvastatin Metabolism and Membrane Transporters in the Small Intestine and Liver of Rats

Membranes ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 233
Author(s):  
Hsien-Tsung Yao ◽  
Ya-Ru Hsu ◽  
Mei-Ling Li
Keyword(s):  
Membrane Protein ◽  
Protein Expression ◽  
Green Tea ◽  
Membrane Transporters ◽  
Male Rats ◽  
Chow Diet ◽  
Beverage Consumption ◽  
Membrane Protein Expression ◽  
P Glycoprotein ◽  
Significant Difference

Green tea (GT) beverages are popular worldwide and may prevent the development of many chronic diseases including cardiovascular disease and cancer. To investigate whether the consumption of a GT beverage causes drug interactions, the effects of GT beverage consumption on atorvastatin metabolism and membrane transporters were evaluated. Male rats were fed a chow diet with tap water or the GT beverage for 3 weeks. Then, the rats were given a single oral dose (10 mg/kg body weight (BW)) of atorvastatin (ATV), and blood was collected at various time points within 6 h. The results show that GT consumption increased the plasma concentrations (AUC0–6h) of ATV (+85%) and 2-OH ATV (+93.3%). GT also increased the 2-OH ATV (+40.9%) and 4-OH ATV (+131.6%) contents in the liver. Decreased cytochrome P450 (CYP) 3A enzyme activity, with no change in P-glycoprotein expression in the intestine, was observed in rats treated with GT. Additionally, GT increased hepatic CYP3A-mediated ATV metabolism and decreased organic anion transporting polypeptides (OATP) 2 membrane protein expression. There was no significant difference in the membrane protein expression of OATP2B1 and P-glycoprotein in the intestine and liver after the GT treatment. The results show that GT consumption may lower hepatic OATP2 and, thus, limit hepatic drug uptake and increase plasma exposure to ATV and 2-OH ATV.

Quantitative Membrane Protein Expression at the Blood–Brain Barrier of Adult and Younger Cynomolgus Monkeys

2011 ◽  
Vol 100 (9) ◽  
pp. 3939-3950 ◽  
Author(s):  
Katsuaki Ito ◽  
Yasuo Uchida ◽  
Sumio Ohtsuki ◽  
Sanshiro Aizawa ◽  
Hirotaka Kawakami ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Expression ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Cynomolgus Monkeys ◽  
Membrane Protein Expression ◽  
Blood Brain
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