Document Detail


Quantum dot targeting with lipoic acid ligase and HaloTag for single-molecule imaging on living cells.
MedLine Citation:
PMID:  23181687     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We present a methodology for targeting quantum dots to specific proteins on living cells in two steps. In the first step, Escherichia coli lipoic acid ligase (LplA) site-specifically attaches 10-bromodecanoic acid onto a 13 amino acid recognition sequence that is genetically fused to a protein of interest. In the second step, quantum dots derivatized with HaloTag, a modified haloalkane dehalogenase, react with the ligated bromodecanoic acid to form a covalent adduct. We found this targeting method to be specific, fast, and fully orthogonal to a previously reported and analogous quantum dot targeting method using E. coli biotin ligase and streptavidin. We used these two methods in combination for two-color quantum dot visualization of different proteins expressed on the same cell or on neighboring cells. Both methods were also used to track single molecules of neurexin, a synaptic adhesion protein, to measure its lateral diffusion in the presence of neuroligin, its trans-synaptic adhesion partner.
Authors:
Daniel S Liu; William S Phipps; Ken H Loh; Mark Howarth; Alice Y Ting
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-12-05
Journal Detail:
Title:  ACS nano     Volume:  6     ISSN:  1936-086X     ISO Abbreviation:  ACS Nano     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-21     Completed Date:  2013-06-11     Revised Date:  2014-01-10    
Medline Journal Info:
Nlm Unique ID:  101313589     Medline TA:  ACS Nano     Country:  United States    
Other Details:
Languages:  eng     Pagination:  11080-7     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Binding Sites
Biocatalysis
Cell Survival
Decanoic Acids / chemistry
Escherichia coli / enzymology
HEK293 Cells
HeLa Cells
Humans
Hydrolases / metabolism*
Ligases / chemistry,  metabolism*
Molecular Imaging / methods*
Peptide Fragments / metabolism
Quantum Dots*
Substrate Specificity
Thioctic Acid / metabolism*
Grant Support
ID/Acronym/Agency:
R01 GM072670/GM/NIGMS NIH HHS; R01 GM072670/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Decanoic Acids; 0/Peptide Fragments; 4G9EDB6V73/decanoic acid; 73Y7P0K73Y/Thioctic Acid; EC 3.-/Hydrolases; EC 3.8.1.5/haloalkane dehalogenase; EC 6.-/Ligases
Comments/Corrections

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


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