Abstract：
:The individual biological activities of many neurotrophic factors and their variants, which are produced by alternative splicing and proteolytic processing, often remain to be characterized. Bacterial protein production combined with protein refolding and purification is a conventional procedure to obtain active neurotrophic factors; however, the procedure is time consuming and appropriate protein refolding in vitro is sometimes unpredictable. Here we examined three distinct cell-free translation systems: reticulocyte lysate, Hela cell lysate and wheat germ extract, which may allow us to produce biologically active factors in a single tube. Taking type-I neuregulin-1 beta3 as an example, we produced neuregulin-1 protein from its mRNAs flanked by Cap nucleotide and/or internal ribosome entry site (IRES) and compared the yields and biological activity of translation products from these systems. The protein yield from IRES+ mRNA was highest in the Hela cell-free system, while background translation was lowest in the wheat germ system. The biological activity of both translation products was modest or negligible, however. Neuregulin-1 protein was produced in reticulocyte lysate at yields of 19 pmol/mL (~500 ng/mL); furthermore, it was potent at phosphorylating ErbB4 receptor and able to bind to heparin sulfate. These results demonstrate that the reticulocyte lysate translation system produces active neurotrophic factors in vitro and is useful for radiolabeling or preliminary assessment of novel neurotrophic factors and their variants.
journal_name
Neurosci Lettjournal_title
Neuroscience lettersauthors
Wang R,Iwakura Y,Araki K,Sotoyama H,Takei N,Nawa Hdoi
10.1016/j.neulet.2011.04.036subject
Has Abstractpub_date
2011-06-22 00:00:00pages
90-3issue
2eissn
0304-3940issn
1872-7972pii
S0304-3940(11)00494-0journal_volume
497pub_type
杂志文章abstract：:The cerebral-buccal interneurons (CBIs) in Aplysia are a group of inter-ganglionic projection neurons that regulate feeding motor programs. In this study, electrophysiological and immunocytological methods were used to identify a previously uncharacterized CBI, designated CBI-11. CBI-11 is a gamma-aminobutyric acid (G...
journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(03)00052-1
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(84)90398-7
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(97)00844-6
更新日期：1997-11-28 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(99)00815-0
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2009.10.092
更新日期：2010-01-14 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(00)01243-x
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2008.02.035
更新日期：2008-04-25 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2009.07.058
更新日期：2010-01-14 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(89)90218-8
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(98)00921-5
更新日期：1999-01-08 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(98)00787-3
更新日期：1998-11-27 00:00:00
abstract：:Saclofen (the direct sulphonic analogue of baclofen) is a competitive antagonist of baclofen at GABAB receptors in guinea pig ileum and rat cortical slices (estimated pA2 = 5.3), at least twice as potent as 2-hydroxy-saclofen (pA2 = 5). A series of related sulphonic analogues also antagonised baclofen in the guinea pi...
journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(89)90824-0
更新日期：1989-12-15 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2014.06.023
更新日期：2014-08-08 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2020.135005
更新日期：2020-05-29 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(91)90368-4
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2014.03.060
更新日期：2014-06-06 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2009.10.051
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journal_title：Neuroscience letters
pub_type： 杂志文章
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更新日期：2000-10-20 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2009.10.090
更新日期：2010-01-04 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(90)90553-l
更新日期：1990-02-05 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/j.neulet.2017.11.033
更新日期：2018-02-05 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(81)90374-8
更新日期：1981-01-20 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章,评审
doi：10.1016/j.neulet.2017.08.005
更新日期：2018-01-10 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(89)90080-3
更新日期：1989-07-31 00:00:00
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journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/s0304-3940(96)13275-4
更新日期：1996-12-27 00:00:00
abstract：:Sympathoexcitatory neurons in the C1 adrenergic area of the rostral ventrolateral medulla (RVL) are tonically inhibited by gamma-aminobutyric acid (GABA). To identify the source of this GABAergic input, the distribution of neurons containing glutamate decarboxylase (GAD) was determined immunocytochemically in rats tre...
journal_title：Neuroscience letters
pub_type： 杂志文章
doi：10.1016/0304-3940(85)90333-7
更新日期：1985-07-04 00:00:00
