Document Detail

Activity-based probes linked with laser-cleavable mass tags for signal amplification in imaging mass spectrometry: analysis of serine hydrolase enzymes in mammalian tissue.
MedLine Citation:
PMID:  22424244     Owner:  NLM     Status:  MEDLINE    
A novel functional imaging mass spectrometry technology is described that utilizes activity-based probes for imaging enzyme active sites in tissue sections. We demonstrate this technology using an activity-based probe (fluorophosphate) that is specific for serine hydrolases. A dendrimer containing multiple mass tags that is attached to the activity-based probe is used to analyze the binding sites of the probe through release and measurement of the mass tags on laser irradiation. A generation 8 poly(amido amine) dendrimer with 1024 amino groups was labeled with an azide group, and then, more than 900 mass tags were attached in order to achieve signal amplification of nearly 3 orders of magnitude. The experimental protocol first involves binding of the activity-based probe containing an alkyne group to serine hydrolases in the tissue section followed by attachment of the dendrimer labeled with mass tags to the bound probe by Click chemistry. On irradiation of the labeled tissue by the laser beam in a raster pattern, the mass tags are liberated and recorded by the mass analyzer; consequently, the ion image of the mass tag reveals the distribution of serine hydrolases in the tissue. This process was shown using rat brain and mouse embryo sections. Targeted imaging has the advantage of providing high spatial resolution and high sensitivity through the use of signal amplification chemistry with high target specificity through the use of an enzyme activity probe.
Junhai Yang; Pierre Chaurand; Jeremy L Norris; Ned A Porter; Richard M Caprioli
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-03-29
Journal Detail:
Title:  Analytical chemistry     Volume:  84     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-04-17     Completed Date:  2012-09-18     Revised Date:  2014-09-22    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3689-95     Citation Subset:  IM    
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MeSH Terms
Brain / enzymology*
Fetus / enzymology*
Mass Spectrometry / methods*
Reference Standards
Serine Proteases / chemistry
Staining and Labeling*
Grant Support
5P41RR031461-02/RR/NCRR NIH HHS; 5R01GM058008/GM/NIGMS NIH HHS; 8 P41 GM103391-02/GM/NIGMS NIH HHS; P41 GM103391/GM/NIGMS NIH HHS; P41 GM103391-02/GM/NIGMS NIH HHS; P41 RR031461/RR/NCRR NIH HHS; P41 RR031461-01/RR/NCRR NIH HHS; R01 GM058008/GM/NIGMS NIH HHS; R01 GM058008-13/GM/NIGMS NIH HHS; R33 CA116123/CA/NCI NIH HHS; R33 CA116123-03/CA/NCI NIH HHS
Reg. No./Substance:
EC 3.4.-/Serine Proteases

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

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