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:  Publisher    
Abstract/OtherAbstract:
We present a methodology for targeting quantum dots to proteins on living cells in two steps. In the first step, E. coli lipoic acid ligase (LplA) site-specifically attaches 10-bromodecanoic acid onto a 13-amino acid peptide that can be 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     Date:  2012-11-26
Journal Detail:
Title:  ACS nano     Volume:  -     ISSN:  1936-086X     ISO Abbreviation:  ACS Nano     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101313589     Medline TA:  ACS Nano     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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