Pyroglutamate aminopeptidase 1 may be an indicator of cellular inflammatory response as revealed using a sensitive long-wavelength fluorescent probe

• Published in: Chemical science (Cambridge) Vol. 7; no. 7; pp. 4694 - 4697
• Main Authors: Gong, Qiuyu, Li, Lihong, Wu, Xiaofeng, Ma, Huimin
• Format: Journal Article
• Language: English
• Published: CAMBRIDGE Royal Soc Chemistry 01.01.2016; Royal Society of Chemistry
• Subjects: Assaying; Cellular; Chemistry; Chemistry, Multidisciplinary; Confocal; Fluorescence; Immunofluorescence; Indicators; Inflammatory response; Physical Sciences; Science & Technology; Stimulation; CELLS; DESIGN; MECHANISMS; ACID; PEPTIDES
• ISSN: 2041-6520; 2041-6539
• DOI: 10.1039/c6sc00951d

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 the immunopotentiators in either RAW264.7 or LO-2 cells is confirmed by measuring tumor necrosis factor alpha (a common inflammatory factor). The above findings indicate that cellular inflammation is accompanied by an increase in PGP-1, and PGP-1 may serve as a new indicator of cellular inflammatory response. Pyroglutamate aminopeptidase 1 may be an indicator of cellular inflammatory response as revealed using a sensitive long-wavelength fluorescent probe.