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rIDE is expressed in insect cells, degrades insulin, and binds to VZV gE.

By Qingxue Li (363724), Mir A. Ali (363725), Kening Wang (291231), Dean Sayre (363726), Frederick G. Hamel (354360), Elizabeth R. Fischer (363727), Robert G. Bennett (354358) and Jeffrey I. Cohen (143240)

Abstract

<p>(A) Structure of HA-tagged rIDE and its promoter in recombinant baculovirus. (B) rIDE was purified, eluted under native conditions, and stained with Coomassie Blue. Soluble rIDE is expressed as a 110 kDa protein in insect cells. (C) HPLC profiles of insulin degradation fragments observed after co-incubation of radiolabeled insulin with rIDE expressed in insect cells, endogenous IDE extracted from rat liver (top panel), or recombinant IDE (6His/Flag-IDE) expressed in eukaryotic cells (bottom panel). (D) Insulin-degrading enzymatic activities of IDE proteins. (E) ELISA assay for binding of rIDE to gEt, gEt lacking the IDE binding domain (gEtΔ32-71), gEt defective in binding to VZV gI (gEtΔ163-208), and a negative control protein (P7.5, vaccinia 7.5 protein). The horizontal dotted line indicates the level of nonspecific binding based on the control protein.</p

Topics: Infectious Diseases, Virology, degrades, binds, vzv
Year: 2013
DOI identifier: 10.1371/journal.pone.0011327.g001
OAI identifier: oai:figshare.com:article/513406
Provided by: FigShare
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