Abstract:
:To investigate the dynamic cellular response to a condition change, selective labeling of the nascent proteome is necessary. Here, we report a method combining click chemistry protein labeling with 2D DIGE. To test the relevance of the method, we compared nascent proteomes of actively growing bacterial cells with that of cells exposed to protein synthesis inhibitor, erythromycin. Cells were incubated with methionine analog, homopropargyl glycin, and their nascent proteome was selectively labeled with monosulfonated neutral Cy3 and Cy5 azides specially synthesized for this purpose. Following fluorescent labeling, the protein samples were mixed and subjected to standard 2D DIGE separation. The method allowed us to reveal a dramatic reduction of newly synthesized proteins upon erythromycin treatment, while the total proteome was not significantly affected. Additionally, several proteins, whose synthesis was resistant to erythromycin, were identified.
journal_name
Proteomicsjournal_title
Proteomicsauthors
Osterman IA,Ustinov AV,Evdokimov DV,Korshun VA,Sergiev PV,Serebryakova MV,Demina IA,Galyamina MA,Govorun VM,Dontsova OAdoi
10.1002/pmic.201200393subject
Has Abstractpub_date
2013-01-01 00:00:00pages
17-21issue
1eissn
1615-9853issn
1615-9861journal_volume
13pub_type
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