Pyroglutamate aminopeptidase 1 may be an indicator of cellular inflammatory response as revealed using a sensitive long-wavelength fluorescent probe
Abstract
Pyroglutamate aminopeptidase 1 (PGP-1) can remove pyroglutamic acid from the N-terminus of a polypeptide, including some important anti-inflammatory proteins. Detecting the change and distribution of cellular PGP-1 in an inflammation process would be helpful to better understand the role of this enzyme. However, no report has been found on this subject, mainly due to the lack of a proper research approach. Herein, we develop such a new method by preparing a sensitive long-wavelength fluorescent probe combined with confocal fluorescence imaging. The probe, consisting of l-pyroglutamic acid and cresyl violet, exhibits high selectivity and sensitivity for PGP-1 under physiological conditions. With this probe, the up-regulation of PGP-1 in LO-2 cells under the stimulation of Freund's incomplete adjuvant and lipopolysaccharide (two main immunopotentiators) is revealed for the first time, and this up-regulation is also observed in typical phagocytic RAW264.7 cells, as evidenced by western blot and inhibition assays. Studies on the distribution of PGP-1 in cells using our probe showed that most PGP-1 is located in the cytoplasm, which is further supported by an immunofluorescence assay. Moreover, the inflammatory response induced by t...Continue Reading
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