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Plos One : Milligram Production and Biological Activity Characterization of the Human Chemokine Receptor Ccr3, Volume 8

By Signoret, Nathalie

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Book Id: WPLBN0003963156
Format Type: PDF eBook :
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Reproduction Date: 2015

Title: Plos One : Milligram Production and Biological Activity Characterization of the Human Chemokine Receptor Ccr3, Volume 8  
Author: Signoret, Nathalie
Volume: Volume 8
Language: English
Subject: Journals, Science, Medical Science
Collections: Periodicals: Journal and Magazine Collection (Contemporary)
Historic
Publication Date:
Publisher: Plos

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Signoret, N. (n.d.). Plos One : Milligram Production and Biological Activity Characterization of the Human Chemokine Receptor Ccr3, Volume 8. Retrieved from http://hawaiilibrary.net/


Description
Description : Human chemokine receptor CCR3 (hCCR3) belongs to the G protein-coupled receptors (GPCRs) superfamily of membrane proteins and plays major roles in allergic diseases and angiogenesis. In order to study the structural and functional mechanism of hCCR3, it is essential to produce pure protein with biological functions on a milligram scale. Here we report the expression of hCCR3 gene in a tetracycline-inducible stable mammalian cell line. A cell clone with high hCCR3 expression was selected from 46 stably transfected cell clones and from this cell line pure hCCR3 on a milligram scale was obtained after two-step purification. Circular dichroism spectrum with a characteristic shape and magnitude for a-helix indicated proper folding of hCCR3 after purification. The biological activity of purified hCCR3 was verified by its high binding affinity with its endogenous ligands CCL11 and CCL24, with KD in the range of 1028 M to 1026 M.

 

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