Abstract
The bifunctional enzyme acylpeptide hydrolase (APEH) is involved in important metabolic processes both as an exopeptidase and as an endopeptidase. Hence, the growing interest in the study of this protein and the need to set up in vitro assays for its characterization. This chapter describes two in vitro assays able to detect the activities of APEH, one for the exopeptidase activity and one for the endopeptidase activity. In particular, these assays have been set up on the two APEH isoforms from Antarctic fish, characterized by a distinct functionality and marked exo- and endopeptidase activities.
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References
Perrier J, Durand A, Giardina T et al (2005) Catabolism of intracellular N-terminal acetylated proteins: involvement of acylpeptide hydrolase and acylase. Biochimie 87:673–685
Shimizu K, Kiuchi Y, Ando K et al (2004) Coordination of oxidized protein hydrolase and the proteasome in the clearance of cytotoxic denatured proteins. Biochem Biophys Res Commun 324:140–146
Gogliettino M, Riccio A, Balestrieri M et al (2014) A novel class of bifunctional acylpeptide hydrolases—potential role in the antioxidant defense systems of the Antarctic fish Trematomus bernacchii. FEBS J 281(1):401–415. https://doi.org/10.1111/febs.12610
Riccio A, Gogliettino M, Palmieri G et al (2015) A new APEH cluster with antioxidant functions in the Antarctic hemoglobinless icefish Chionodraco hamatus. PLoS One 10(5):e0125594. https://doi.org/10.1371/journal.pone.0125594. PMID: 25946123; PMCID: PMC4422685
Brand L, Johnson ML (eds) (1997) “Fluorescence Spectroscopy” methods in enzymology, vol 278. Academic, San Diego
Bisswanger H (2011) Practical enzymology. Wiley-VCH Verlag. https://doi.org/10.1002/9783527659227
Vandooren J, Geurts N, Martens E et al (2013) Zymography methods for visualizing hydrolytic enzymes. Nat Methods 10:211–220. https://doi.org/10.1038/nmeth.2371
Fujino T, Kojima M, Beppu M et al (2000) Identification of the cleavage sites of oxidized protein that are susceptible to oxidized protein hydrolase (OPH) in the primary and tertiary structures of the protein. J Biochem 127(6):1087–1093. https://doi.org/10.1093/oxfordjournals.jbchem.a022702
Fujino T, Ishikawa T, Inoue M et al (1998) Characterization of membrane-bound serine protease related to degradation of oxidatively damaged erythrocyte membrane proteins. Biochim Biophys Acta 1374:47–55
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Fusco, C., Agrillo, B., Gogliettino, M., Palmieri, G., Cocca, E. (2022). In Vitro Assays for the Bifunctional Acylpeptide Hydrolase (APEH) Enzyme from Antarctic Fish. In: Verde, C., Giordano, D. (eds) Marine Genomics. Methods in Molecular Biology, vol 2498. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2313-8_25
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DOI: https://doi.org/10.1007/978-1-0716-2313-8_25
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