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Selective Homogeneous Assay for Circulating Endopeptidase Fibroblast Activation Protein (FAP)
- Source :
- Scientific Reports, Vol 7, Iss 1, Pp 1-12 (2017), Scientific Reports
- Publication Year :
- 2017
- Publisher :
- Springer Science and Business Media LLC, 2017.
-
Abstract
- Fibroblast Activation Protein (FAP) is a membrane-bound serine protease whose expression is often elevated in activated fibroblasts associated with tissue remodeling in various common diseases such as cancer, arthritis and fibrosis. Like the closely related dipeptidyl peptidase DPPIV, the extracellular domain of FAP can be released into circulation as a functional enzyme, and limited studies suggest that the circulating level of FAP correlates with the degree of tissue fibrosis. Here we describe a novel homogeneous fluorescence intensity assay for circulating FAP activity based on a recently identified natural substrate, FGF21. This assay is unique in that it can effectively distinguish endopeptidase activity of FAP from that of other related enzymes such as prolyl endopeptidase (PREP) and was validated using Fap-deficient mice. Structural modeling was used to elucidate the mechanistic basis for the observed specificity in substrate recognition by FAP, but not by DPPIV or PREP. Finally, the assay was used to detect elevated FAP activity in human patients diagnosed with liver cirrhosis and to determine the effectiveness of a chemical inhibitor for FAP in mice. We propose that the assay presented here could thus be utilized for diagnosis of FAP-related pathologies and for the therapeutic development of FAP inhibitors.
- Subjects :
- Liver Cirrhosis
0301 basic medicine
congenital, hereditary, and neonatal diseases and abnormalities
lcsh:Medicine
Gene Expression Regulation, Enzymologic
Article
Dipeptidyl peptidase
Substrate Specificity
Mice
03 medical and health sciences
Endopeptidase activity
Fibroblast activation protein, alpha
Prolyl endopeptidase
Fibrosis
Endopeptidases
medicine
Animals
Humans
lcsh:Science
neoplasms
Dipeptidyl peptidase-4
Serine protease
Multidisciplinary
biology
Chemistry
Serine Endopeptidases
lcsh:R
Membrane Proteins
Fibroblasts
medicine.disease
Molecular biology
digestive system diseases
Endopeptidase
Fibroblast Growth Factors
030104 developmental biology
Gelatinases
biology.protein
lcsh:Q
Prolyl Oligopeptidases
medicine.drug
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....93dc3ba0acd119f5bfbac60538e2ad60