Document Detail


Fluorescence labeling of a cytokine with desthiobiotin-tagged fluorescent puromycin.
MedLine Citation:
PMID:  18397775     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fluorescence labeling of a cytokine at a specific site is required for observing cytokine-receptor interactions in living cells at the single-molecule level. Here, we demonstrated the C-terminus-specific fluorescence labeling of histidine-tagged thrombopoietin (TPO), a ligand for Mpl, with desthiobiotin-tagged fluorescent puromycin. Fluorescent TPO, purified by tandem affinity purification, stimulated the proliferation of Mpl-expressing cells. Within 10 min of its addition, fluorescent TPO was found to be diffusely distributed on the cell membranes of Mpl-expressing cells, and gradually accumulated to form fluorescent spots. This method is applicable for studying the spatial and temporal distributions of cytokines in individual living cells.
Authors:
Akihiko Sakamoto; Mai Yamagishi; Takafumi Watanabe; Youichi Aizawa; Takashi Kato; Takashi Funatsu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of bioscience and bioengineering     Volume:  105     ISSN:  1389-1723     ISO Abbreviation:  J. Biosci. Bioeng.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-04-09     Completed Date:  2008-08-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100888800     Medline TA:  J Biosci Bioeng     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  238-42     Citation Subset:  IM    
Affiliation:
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biotin / analogs & derivatives*,  chemistry
Cell Line
Fluorescent Dyes / chemistry*
Mice
Puromycin / chemistry*
Thrombopoietin / chemistry,  metabolism*
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 15720-25-9/desthiobiotin; 53-79-2/Puromycin; 58-85-5/Biotin; 9014-42-0/Thrombopoietin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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