scholarly journals Expression and Purification of Membrane Proteins in Saccharomyces cerevisiae

2020 ◽  
pp. 47-61
Author(s):  
Martin S. King ◽  
Edmund R. S. Kunji
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Proteins ◽  
Expression And Purification

scholarly journals Prohibitins Regulate Membrane Protein Degradation by the m-AAA Protease in Mitochondria

1999 ◽  
Vol 19 (5) ◽  
pp. 3435-3442 ◽  
Author(s):  
Gregor Steglich ◽  
Walter Neupert ◽  
Thomas Langer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Growth ◽  
Membrane Proteins ◽  
Eukaryotic Cells ◽  
Mitochondrial Morphology ◽  
Regulatory Effect ◽  
Inner Membrane ◽  
Functional Activities ◽  
Native Molecular Mass ◽  
Affect Cell Growth

ABSTRACT Prohibitins comprise a protein family in eukaryotic cells with potential roles in senescence and tumor suppression. Phb1p and Phb2p, members of the prohibitin family in Saccharomyces cerevisiae, have been implicated in the regulation of the replicative life span of the cells and in the maintenance of mitochondrial morphology. The functional activities of these proteins, however, have not been elucidated. We demonstrate here that prohibitins regulate the turnover of membrane proteins by the m-AAA protease, a conserved ATP-dependent protease in the inner membrane of mitochondria. The m-AAA protease is composed of the homologous subunits Yta10p (Afg3p) and Yta12p (Rca1p). Deletion ofPHB1 or PHB2 impairs growth of Δyta10 or Δyta12 cells but does not affect cell growth in the presence of the m-AAA protease. A prohibitin complex with a native molecular mass of approximately 2 MDa containing Phb1p and Phb2p forms a supercomplex with them-AAA protease. Proteolysis of nonassembled inner membrane proteins by the m-AAA protease is accelerated in mitochondria lacking Phb1p or Phb2p, indicating a negative regulatory effect of prohibitins on m-AAA protease activity. These results functionally link members of two conserved protein families in eukaryotes to the degradation of membrane proteins in mitochondria.

Tvp38, Tvp23, Tvp18 and Tvp15: Novel membrane proteins in the Tlg2-containing Golgi/endosome compartments of Saccharomyces cerevisiae

2007 ◽  
Vol 313 (4) ◽  
pp. 688-697 ◽  
Author(s):  
Hironori Inadome ◽  
Yoichi Noda ◽  
Yurika Kamimura ◽  
Hiroyuki Adachi ◽  
Koji Yoda
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Proteins

scholarly journals Cloning, expression and purification of recombinant Slx4 from Saccharomyces cerevisiae (539.3)

2014 ◽  
Vol 28 (S1) ◽  
Author(s):  
Angel Diaz Martinez ◽  
Jose Renato Cussiol ◽  
Marcus Smolka
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Expression And Purification

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 Fmp30, Mdm31, and Mdm32 Function in Ups1-Independent Cardiolipin Accumulation Under Low Phosphatidylethanolamine Conditions

Contact ◽  
2018 ◽  
Vol 1 ◽  
pp. 251525641876404
Author(s):  
Non Miyata ◽  
Osamu Kuge
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Proteins ◽  
Phosphatidic Acid ◽  
Yeast Saccharomyces Cerevisiae ◽  
Inner Membrane ◽  
Mitochondrial Inner Membrane ◽  
Per Se

Maintenance of the cardiolipin (CL) level largely depends on Ups1-Mdm35 complex-mediated intramitochondrial phosphatidic acid transfer. In addition, the presence of an alternative CL accumulation pathway has been suggested in the yeast Saccharomyces cerevisiae. This pathway is independent of the Ups1-Mdm35 complex and stimulated by loss of Ups2, which forms a complex with Mdm35 and mediates intramitochondrial transfer of phosphatidylserine for phosphatidylethanolamine synthesis. Recently, we found that the alternative CL accumulation pathway is enhanced by a lowered phosphatidylethanolamine level, not by loss of Ups2 per se, and depends on three mitochondrial inner membrane proteins, Fmp30, Mdm31, and Mdm32.

scholarly journals Heterologous Expression and Purification Systems for Structural Proteomics of Mammalian Membrane Proteins

2002 ◽  
Vol 3 (6) ◽  
pp. 511-517 ◽  
Author(s):  
Isabelle Mus-Veteau
Keyword(s):  
Membrane Proteins ◽  
Heterologous Expression ◽  
Large Scale ◽  
Structural Information ◽  
Structural Data ◽  
Structural Proteomics ◽  
Scale Production ◽  
Expression And Purification ◽  
Large Scale Production ◽  
Heterologous Expression Systems

Membrane proteins (MPs) are responsible for the interface between the exterior and the interior of the cell. These proteins are implicated in numerous diseases, such as cancer, cystic fibrosis, epilepsy, hyperinsulinism, heart failure, hypertension and Alzheimer's disease. However, studies on these disorders are hampered by a lack of structural information about the proteins involved. Structural analysis requires large quantities of pure and active proteins. The majority of medically and pharmaceutically relevant MPs are present in tissues at very low concentration, which makes heterologous expression in large-scale production-adapted cells a prerequisite for structural studies. Obtaining mammalian MP structural data depends on the development of methods that allow the production of large quantities of MPs. This review focuses on the different heterologous expression systems, and the purification strategies, used to produce large amounts of pure mammalian MPs for structural proteomics.

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