From gene to protein: a review of new and enabling technologies for multi-parallel protein expression

Hunt, I. V.

doi:10.1016/j.pep.2004.10.018

Cited by 167 publications

(109 citation statements)

References 94 publications

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“…Structural Modifications, Purification, and Expression Systems Do Not Affect ProNGF Biological Activity-Amino acid substitutions may disturb the ability of a molecule to bind to its receptor (44), whereas the presence of polyhistidine COOHterminal extensions may modify the active site of a protein (45), disturb protein structure (46,47), or alter biochemical properties such as dimerization/oligomerization (48). Here we show that proNGF carrying four amino acid substitutions and a COOH-terminal histidine tag (30), carrying two amino acid substitutions and no tag (33), or carrying six amino acid substitutions and no tag (31) promoted neurite outgrowth or elicited TrkA and MAPK phosphorylation with activity similar to proNGF(RϪ1G) with only one amino acid substitution and no tag (32).…”

Section: Discussionmentioning

confidence: 99%

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Biological Activity of Nerve Growth Factor Precursor Is Dependent upon Relative Levels of Its Receptors

Masoudi¹,

Ioannou²,

Coughlin

et al. 2009

Journal of Biological Chemistry

117

109

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Nerve growth factor (NGF) is produced as a precursor called pro-nerve growth factor (proNGF), which is secreted by many tissues and is the predominant form of NGF in the central nervous system. In Alzheimer disease brain, cholinergic neurons degenerate and can no longer transport NGF as efficiently, leading to an increase in untransported NGF in the target tissue. The protein that accumulates in the target tissue is proNGF, not the mature form. The role of this precursor is controversial, and both neurotrophic and apoptotic activities have been reported for recombinant proNGFs. Differences in the protein structures, protein expression systems, methods used for protein purification, and methods used for bioassay may affect the activity of these proteins. Here, we show that proNGF is neurotrophic regardless of mutations or tags, and no matter how it is purified or in which system it is expressed. However, although proNGF is neurotrophic under our assay conditions for primary sympathetic neurons and for pheochromocytoma (PC12) cells, it is apoptotic for unprimed PC12 cells when they are deprived of serum. The ratio of tropomyosin-related kinase A to p75 neurotrophin receptor is low in unprimed PC12 cells compared with primed PC12 cells and sympathetic neurons, altering the balance of proNGFinduced signaling to favor apoptosis. We conclude that the relative level of proNGF receptors determines whether this precursor exhibits neurotrophic or apoptotic activity.

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Section: Discussionmentioning

confidence: 99%

“…Nickel column chromatography can cause oxidation and promote proteolysis, change protein properties, or cause protein aggregation (46,47,49). Here we show that purification over a nickel column does not alter the neurotrophic activity of proNGF.…”

Section: Discussionmentioning

confidence: 99%

Biological Activity of Nerve Growth Factor Precursor Is Dependent upon Relative Levels of Its Receptors

Masoudi¹,

Ioannou²,

Coughlin

et al. 2009

Journal of Biological Chemistry

117

109

View full text Add to dashboard Cite

show abstract

“…E. coli BL21 is an example of the most common host and it has been proven outstanding in application for standard recombinant expression. It can grow efficiently in minimal media as nonpathogenic bacterium that cannot survive to cause diseases in host tissues [9,11,15,16] .…”

Section: Modification Of E Coli Host Strainmentioning

confidence: 99%

“…The increase of expression and activity of lower temperatures growth is associated with increased expression of chaperones in E. coli. Therefore, growth at a temperature range of (15)(16)(17)(18)(19)(20)(21)(22)(23) 曟, could also lead to a significant reduction of expressed protein degradation [4,18,19] .…”

Section: Expression At Lower Temperaturesmentioning

confidence: 99%

Strategies for production of active eukaryotic proteins in bacterial expression system

Khow

Suntrarachun

2012

Asian Pacific Journal of Tropical Biomedicine

114

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“…La présence de corps d'inclusion qui rendent difficile la purification, et la production d'endotoxines complètent cette liste de défauts. En conséquence, la production de protéines recombinantes complexes requiert l'usage de systèmes d'expression eucaryotes [2].…”

Section: Les Différents Systèmes D'expressionunclassified

Les microalgues

et al. 2008

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From gene to protein: a review of new and enabling technologies for multi-parallel protein expression

Cited by 167 publications

References 94 publications

Biological Activity of Nerve Growth Factor Precursor Is Dependent upon Relative Levels of Its Receptors

Biological Activity of Nerve Growth Factor Precursor Is Dependent upon Relative Levels of Its Receptors

Strategies for production of active eukaryotic proteins in bacterial expression system

Les microalgues

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