The purification and characterisation of novel dipeptidyl peptidase IV-like activity from bovine serum

Buckley, Seamus J; Collins, Patrick J.; O’Connor, Brendan F.

doi:10.1016/j.biocel.2003.02.001

Cited by 13 publications

(15 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The data showed that the metallopeptidase inhibitor phenanthroline completely inhibited Gly-Pro-AMC and l -Leu-AMC cleavage (Table 2). Although classified as serine protease, dipeptidyl peptidase 4 is not much sensible to PMSF and its inhibition by phenanthrolines was already noticed to a DPP4 like from bovine serum [50]. The hydrolysis inhibition of l -Leu-AMC by phenanthroline together with the optimal pH results (pH 7.0) suggested the presence of metallopeptidases.…”

Section: Resultsmentioning

confidence: 99%

The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

et al. 2014

View full text Add to dashboard Cite

Peptidases are ubiquitous enzymes involved in diverse biological processes. Fragments from bioactive peptides have been found in skin secretions from frogs, and their presence suggests processing by peptidases. Thus, the aim of this work was to characterize the peptidase activity present in the skin secretion of Leptodactylus labyrinthicus. Zymography revealed the presence of three bands of gelatinase activity of approximately 60 kDa, 66 kDa, and 80 kDa, which the first two were calcium-dependent. These three bands were inhibited either by ethylenediaminetetraacetic acid (EDTA) and phenathroline; thus, they were characterized as metallopeptidases. Furthermore, the proteolytic enzymes identified were active only at pH 6.0–10.0, and their activity increased in the presence of CHAPS or NaCl. Experiments with fluorogenic substrates incubated with skin secretions identified aminopeptidase activity, with cleavage after leucine, proline, and alanine residues. This activity was directly proportional to the protein concentration, and it was inhibited in the presence of metallo and serine peptidase inhibitors. Besides, the optimal pH for substrate cleavage was determined to be 7.0–8.0. The results of the in gel activity assay showed that all substrates were hydrolyzed by a 45 kDa peptidase. Gly-Pro-AMC was also cleaved by a peptidase greater than 97 kDa. The data suggest the presence of dipeptidyl peptidases (DPPs) and metallopeptidases; however, further research is necessary. In conclusion, our work will help to elucidate the implication of these enzymatic activities in the processing of the bioactive peptides present in frog venom, expanding the knowledge of amphibian biology.

show abstract

Section: Resultsmentioning

confidence: 99%

The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Processing of many bioactive peptides (substance P) and circulating peptide hormones (growth hormone-releasing hormone GRH) takes place in the blood circulation by DPP IV. Therefore, DPP IV needs to be active and stable at this pH in order to process these bioactive peptides [2,5,7].…”

Section: Discussionmentioning

confidence: 99%

“…Enzyme Preparation Dipeptidyl Peptidase IV-like activity was purified from whole bovine serum to near homogeneity (specific activity 1.1 U/mg) using hydrophobic interaction (Phenyl Sepharose 4B), gel filtration (Sephacryl S-300) and anion-exchange (Q-Sepharose) chromatographies in buffers based on 50mM HEPES pH 8.0, as described by Buckley et al [7].…”

Section: Methodsmentioning

confidence: 99%

“…In vivo, DPP IV is ubiquitous, occurring in both membrane-bound and soluble forms [6] and has diverse roles in various cell types [7]. It participates in the post-translational processing of chemokines (such as RANTES) and in the inactivation of neuropeptides (such as substance P) [4,5,8].…”

Section: Introductionmentioning

confidence: 99%

“…refs. [7]). Here we show that bovine serum DPP IV-like peptidase is very stable to water-miscible organic solvents and possesses good thermal stability characteristics.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum

Ruth

Buckley

O’Connor

et al. 2007

Enzyme and Microbial Technology

View full text Add to dashboard Cite

show abstract

Iterative design of emetine‐based prodrug targeting fibroblast activation protein (FAP) and dipeptidyl peptidase IV DPPIV using a tandem enzymatic activation strategy

et al. 2016

View full text Add to dashboard Cite

This FAP activated prodrug of cytotoxic agent emetine further shows the crucial role of the N-2' position of emetine in controlling its cytotoxicity. Significantly reduced toxicity observed in the PrEC cell line in the absence of FAP and DPPIV shows that prodrug 11 could be systemically delivered to regions of metastatic prostate cancer or other solid tumor for activation by cancer selective enzymes within the cancer microenvironment, such as FAP that is overexpressed by the carcinoma-associated fibroblasts. The two-step tandem enzymatic activation of prodrug 11 by FAP and DPPIV is a strategy for overcoming steric hindrance.

show abstract

The purification and characterisation of novel dipeptidyl peptidase IV-like activity from bovine serum

Cited by 13 publications

References 33 publications

The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum

Iterative design of emetine‐based prodrug targeting fibroblast activation protein (FAP) and dipeptidyl peptidase IV DPPIV using a tandem enzymatic activation strategy

Contact Info

Product

Resources

About