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.

Expression and Purification of Serine Protease Inhibitor, OH-TCI, in Escherichia coli as a Thioredoxin Fusion Protein

2014 ◽  
Vol 926-930 ◽  
pp. 1100-1103
Author(s):  
Tong Yi Sun
Keyword(s):  
Large Scale ◽  
Blood Product ◽  
Serine Proteinase ◽  
Serine Protease Inhibitor ◽  
Blood Product Transfusion ◽  
Scale Production ◽  
Serine Proteinase Inhibitor ◽  
Expression And Purification ◽  
Preclinical Evaluation ◽  
Large Scale Production

The OH-TCI was the first report of Kunitz/BPTI serine proteinase inhibitor from snake venom with strong inhibitory activity against trypsin and chymotrypsin. Thus it is theoretically attractive in ameliorating the effects of acute pancreatitis and reducing allogeneic blood product transfusion during cardiac surgery. To accomplish preclinical evaluation with OH-TCI, large-scale production of OH-TCI is produced in Escherichia coli. The optimized OH-TCI codons were cloned into pET32a (+). OH-TCI expressed as Trx tag is solubility as fusion bodies. The amount of the purified Trx-OH-TCI from 1 liter culture was roughly 200 mg. The inhibitor constants (Ki) of Trx-OH-TCI, similar to the native OH-TCI, were 2.67×10-7 and 3.60×10-7 M M for trypsin and chymotrypsin, respectively.

Large-scale production of functional membrane proteins

2008 ◽  
Vol 65 (11) ◽  
pp. 1729-1755 ◽  
Author(s):  
F. Junge ◽  
B. Schneider ◽  
S. Reckel ◽  
D. Schwarz ◽  
V. Dötsch ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Large Scale ◽  
Scale Production ◽  
Large Scale Production ◽  
Functional Membrane

I. S. A. Baud. Forms of Production and Women's Labour: Gender Aspects of Industrialisation in India and Mexico. New Delhi: Sage Publications. 1992. 321 pp.Hardbound. Price: Indian Rs 295.00.

1993 ◽  
Vol 32 (1) ◽  
pp. 129-131
Author(s):  
Naureen Talha
Keyword(s):  
Large Scale ◽  
Indian Society ◽  
New Delhi ◽  
Scale Production ◽  
Female Labour ◽  
Self Employment ◽  
Commodity Production ◽  
Large Scale Production ◽  
Mexican Society ◽  
Small Production

The literature on female labour in Third World countries has become quite extensive. India, being comparatively more advanced industrially, and in view of its size and population, presents a pictures of multiplicity of problems which face the female labour market. However, the author has also included Mexico in this analytical study. It is interesting to see the characteristics of developing industrialisation in two different societies: the Indian society, which is conservative, and the Mexican society, which is progressive. In the first chapter of the book, the author explains that he is not concerned with the process of industrialisation and female labour employed at different levels of work, but that he is interested in forms of production and women's employment in large-scale production, petty commodity production, marginal small production, and self-employment in the informal sector. It is only by analysis of these forms that the picture of females having a lower status is understood in its social and political setting.

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