Gold Nanoparticle-Based Colorimetric and Electrochemical Methods for Dipeptidyl Peptidase-IV Activity Assay and Inhibitor Screening
نویسندگان
چکیده
We presented the colorimetric and electrochemical methods for determination of the dipeptidyl peptidase-IV (DPP-IV) activity and screening of its inhibitor using gold nanoparticle (AuNP) as the probe. In the colorimetric assay, the substrate peptide with a sequence of Arg-Pro-Arg induced the aggregation and color change of AuNPs, whereas cleavage of the peptide by DPP-IV prevented the aggregation of AuNPs. Furthermore, the aggregation of AuNPs in the solution was easily initiated on a solid/liquid (electrode/electrolyte) surface, which induced a decrease in the electron-transfer resistance. However, once the peptide was clipped by DPP-IV, the assembly of AuNPs on electrode surface was prevented. Consequently, a higher electron-transfer resistance was observed. The colorimetric and electrochemical assays allowed for the determination of DPP-IV with the detection limits of 70 μU/mL and 0.55 μU/mL, respectively. Meanwhile, the proposed methods were used to determine DPP-IV inhibitor with satisfactory results. Both the colorimetric and electrochemical methods are simple, rapid and sufficiently sensitive for DPP-IV activity assay and inhibitor screening. The results also demonstrated that the AuNP-based colorimetric assay could be converted into an enhanced surface tethered electrochemical assay with improving sensitivity. The simple detection principle may be extended to the design of other peptidases biosensors with easy manipulation procedures.
منابع مشابه
Activity of Dipeptidyl Peptidase-IV/CD26 and Aminopeptidase N/CD13 in Secretome of Mesenchymal Stem Cells after Treatment with LPS and PMA
Background: Emerging evidence suggests that secretome of mesenchymal stem cells has many anti-inflammatory and regenerative properties, which makes it a suitable candidate for the treatment of autoimmune and degenerative diseases. Dipeptidyl Peptidase-IV (DPP-IV)/CD26 and Aminopeptidase N (APN)/CD13 are ubiquitous ecto-enzymes which can digest various substrates including some ...
متن کاملAssessing Stability of Gold Nanoparticles in Presence of Two Enzymes, RNaseA and RNaseH, Using Colorimetric Detection
Gold nanoparticle-based diagnostic methods have attracted much attention due to their simplicity, high sensitivity and low cost. These methods are mostly used for early and efficient detection of various pathogenic factors and assessment of gene expression and nanoparticles stability. In this study, melon plants were cultured in vivo and then, RNA was extracted from leaf explants. A 40 bp probe...
متن کاملDipeptidyl peptidase-4 inhibitor ameliorates status epilepticus seizures and cognitive disturbances in a rat model of temporal lobe epilepsy
Background and Objective: In temporal lobe epilepsy (TLE), recurrent seizures accompany with cognitive deficit. In some patients, the current medications cannot provide satisfactory control of seizures, therefore new drugs that act through different mechanisms are required. In the present study, the useful effect of dipeptidyl peptidase-4 inhibitor was evaluated in experimental model of tempora...
متن کاملDipeptidyl peptidase IV in angiotensin-converting enzyme inhibitor associated angioedema.
Angioedema is a potentially life-threatening adverse effect of angiotensin-converting enzyme inhibitors. Bradykinin and substance P, substrates of angiotensin-converting enzyme, increase vascular permeability and cause tissue edema in animals. Studies indicate that amino-terminal degradation of these peptides, by aminopeptidase P and dipeptidyl peptidase IV, may be impaired in individuals with ...
متن کاملCharacterization and Evaluation of the Activity of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis
Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infec...
متن کامل