Application of heptapeptides containing D‐amino acid residues immobilized to magnetic particles and Sepharose for the study of binding properties of gastric aspartic proteases

Abstract

Synthetic heptapeptides containing D-amino acid residues and differing in the content of L-phenylalanine and L-tyrosine residues and their position (Val-D-Leu-Pro-Tyr-Phe-Val-D-Leu, Val-D-Leu-Pro-Tyr-Tyr-Val-D-Leu, Val-D-Leu-Pro-Phe-Tyr-Val-D-Leu) were immobilized to two types of carriers: glyoxal-activated magnetic agarose particles and CNBr-activated Sepharose. In both cases, peptides were immobilized via their terminal amino group. Immobilized peptides were used for the study of binding properties of two gastric aspartic proteases (porcine pepsin A and rat pepsin C). Porcine pepsin A was adsorbed to all studied peptide-modified magnetic carriers, while rat pepsin C interacted with immobilized ligands only slightly. Similar results were obtained in affinity chromatographic experiments using heptapeptides immobilized to Sepharose.

Magnetic chelating nanoprobes for enrichment and selective recovery of metalloproteases from human saliva

“…Scientic reports regarding the development and applications of magnetic nanoprobes for the enrichment of proteases are scarce. To the best of our knowledge, only Filuszová and collaborators 29,30 developed magnetic bionanoprobes for the selective enrichment of proteases, namely, aspartic proteases. It should be noted, however, that the work reported was not related to the testing of biological samples.…”

Aptamer‐affinity column clean‐up coupled with ultra high performance liquid chromatography and fluorescence detection for the rapid determination of ochratoxin A in ginger powder

“…However, CNBr possesses high toxicity to human health and unfriendliness to the environment. Furthermore, CNBr‐coated immobilization is time consuming, which may disable the bioactivity of ligands . Conversely, immobilization activated by N ‐hydroxysuccinimide (NHS) is a rapid, simple and efficient coupling strategy that is widely appropriate for micro‐ and macromolecules immobilization.…”

Strategies for analysis of isomeric peptides

“…It should be noted that reliance on aminopeptidase M alone carries a risk for false‐positives in terms of DAACP detection, because the presence of proline in the peptide will also inhibit further digestion, analogously with a d ‐amino acid residue. Furthermore, due to their resistance to degradation, DAACPs have also been used in an affinity‐based procedure for separation of digestive enzymes such as porcines .…”