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Review
. 2024 Jun:689:115506.
doi: 10.1016/j.ab.2024.115506. Epub 2024 Mar 7.

Prolidase activity assays. A survey of the reported literature methodologies

Serena Fiorito  1 , Salvatore Genovese  2 , Francesco Epifano  3 , Chiara Collevecchio  1
Affiliations
Free article
Review

Prolidase activity assays. A survey of the reported literature methodologies

Serena Fiorito et al. Anal Biochem. 2024 Jun.
Free article

Abstract

Prolidase (EC.3.4.13.9) is a dipeptidase known nowadays to play a pivotal role in several physiological and pathological processes. More in particular, this enzyme is involved in the cleavage of proline- and hydroxyproline-containing dipeptides (imidodipeptides), thus finely regulating the homeostasis of free proline and hydroxyproline. Abnormally high or low levels of prolidase have been found in numerous acute and chronic syndromes affecting humans (chronic liver fibrosis, viral and acute hepatitis, cancer, neurological disorders, inflammation, skin diseases, intellectual disability, respiratory infection, and others) for which the content of proline is well recognized as a clinical marker. As a consequence, the accurate analytical determination of prolidase activity is of greatly significant importance in clinical diagnosis and therapy. Apart from the Chinard's assay, some other more sensitive and well validated methodologies have been published. These include colorimetric and spectrophotometric determinations of free proline produced by enzymatic reactions, capillary electrophoresis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, electrochemoluminescence, thin layer chromatography, and HPLC. The aim of this comprehensive review is to make a detailed survey of the in so far reported analytical techniques, highlighting their general features, as well as their advantages and possible drawbacks, providing in the meantime suggestions to stimulate further research in this intriguing field.

Keywords: Chinard's reaction; Collagen; Enzymatic assays; Imido dipeptides; Prolidase; Proline.

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Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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