Abstract
AbstractThe lysosomal endoprotease legumain (asparaginyl endoprotease) has been proposed as a putative biomarker in prostate tumours, in which the enzyme is markedly overexpressed. Overexpression, coupled with highly selective specificity for cleavage of substrates at the C-terminus of asparagine (Asn) residues, make legumain an attractive biochemical target for potential diagnosis, prognosis and treatment. We report the design, synthesis, characterisation and preliminary evaluation of a new rhodamine-B (Rho-B)-labelled legumain peptide substrate probe5[Rho-Pro-Ala-Asn-PEG-AQ(4-OH)] and its selective targeting to lysosomes in PC3 prostate cancer cells. Probe5was efficiently activated by recombinant human legumain to afford the high quantum yield reporter fluorophore tripeptide4b(Rho-Pro-Ala-Asn-OH) with concomitant release of intense fluorescence. Furthermore, probe5was activated upon incubation with homogenates derived from fresh-frozen tissue material of prostatectomy specimens. Probe5represents a new viable biochemical tool for probing the activity of legumain with the potential to be used in ex vivo diagnostics in the cancer pathology laboratory.
Publisher
Springer Science and Business Media LLC
Subject
Drug Discovery,Molecular Medicine,Biochemistry,Bioengineering,Analytical Chemistry
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