Size-Dependent Cellular Uptake of RGD Peptides

Monomeric RGD peptides show unspecific fluid-phase uptake in cells, whereas multimeric RGD peptides are thought to be internalized by integrin-mediated endocytosis. However, a potential correlation between uptake mechanism and molecular mass has been neglected so far. A dual derivatization of peptide c(RGDw(7Br)K) was performed to investigate this. A fluorescent probe was installed by chemoselective Suzuki–Miyaura cross-coupling of the 7-bromotryptophan and a poly(ethylene glycol) (PEG) linker was attached to the lysine residue. Flow cytometry and live cell imaging confirmed unspecific uptake of the small, non-PEGylated peptide, whereas the PEG₅₀₀₀ peptide conjugate unveiled a selective internalization by M21 cells overexpressing α_vβ₃ and no uptake in α_v-deficient M21L cells.     

Cellular Disulfide Bond Formation in Bioactive Peptides and Proteins

Bioactive peptides play important roles in metabolic regulation and modulation and many are used as therapeutics. These peptides often possess disulfide bonds, which are important for their structure, function and stability. A systematic network of enzymes—a disulfide bond generating enzyme, a disulfide bond donor enzyme and a redox cofactor—that function inside the cell dictates the formation and maintenance of disulfide bonds. The main pathways that catalyze disulfide bond formation in peptides and proteins in prokaryotes and eukaryotes are remarkably similar and share several mechanistic features. This review summarizes the formation of disulfide bonds in peptides and proteins by cellular and recombinant machinery.     

Selective separation and characterisation of dual ACE and DPP-IV inhibitory peptides from rainbow trout (Oncorhynchus mykiss) protein hydrolysates

Microwave pretreatment and hydrolysis were applied to rainbow trout (Oncorhynchus mykiss) by-products to produce bioactive peptides with dual in vitro angiotensin-I converting enzyme (ACE) and dipeptidyl-peptidase IV (DPP-IV) inhibitory activities. Peptides were fractionated using the single step electrodialysis with ultrafiltration membrane (EDUF). Concentration of cationic peptides (CP) increased in the recovery solution, reaching 125 μg mL⁻¹ after a 4-h treatment with migration rate of 15.68 ± 2.98 g m⁻² h. CP fractions displayed ACE and DPP-IV I inhibitory properties, with IC₅₀ values of 0.0036 mg mL⁻¹ and 1.23 mg mL⁻¹ respectively. The bioactivity was attributed to the low molecular weight peptides (300–500 Da) recovered. CP exhibited non-competitive inhibition patterns for ACE and DPP-IV, which were dose dependent. These results showed that bioactive peptides can successfully be separated from complex hydrolysate mixtures by EDUF. The fractionated peptides can serve as potential functional food ingredients or nutraceuticals for the management of hypertension and diabetes.     

Enhancing the Cellular Delivery of Nanoparticles Using Lipo‐Oligoarginine Peptides

Nanoparticles have great potential as nanotherapeutics, delivery vectors, and molecular imaging agents due to their flexible properties. Although intracellular and nuclear delivery of nanoparticles is desirable for therapeutic applications, it remains a challenge. Cell penetrating peptides (CPPs) are a powerful tool for the intracellular delivery of various cargoes. Here it is reported that functionalization of nanoparticles with a myristoylated oligoarginine CPP promotes cellular uptake without increased toxicity. It is evident that the myristoylated CPP is much more effective in transporting nanoparticles than the unmodified CPPs.     

Capillary electrophoretic profiling of tryptic digests of water soluble proteins from Bacillus thuringiensis-transgenic and non-transgenic maize species

Capillary zone electrophoresis (CZE) has been applied to separation and characterisation of enzymatic (tryptic) hydrolysates of water-soluble proteins from Bacillus thuringiensis (Bt)-transgenic (Aristis-Bt) and two native non-transgenic (Aristis and Coventry) maize varieties. Water-soluble proteins were extracted from the flour of these maize species and digested by bovine pancreatic trypsin immobilised on agarose gel in 100 mM ammonium hydrocarbonate buffer, pH 7.9. The yielded tryptic digests of proteins were analysed by CZE in four acidic background electrolytes (BGEs) (100 mM H₃PO₄, 50 mM Tris, pH 2.25; 500 mM acetic acid, pH 2.54; 200 mM formic acid, 200 mM acetic acid, pH 2.05; and 200 mM iminodiacetic acid, pH 2.26) using a lab-made CZE apparatus equipped with bare fused silica capillary and UV-absorption detector operating at 206 nm. Among the tested BGEs, the best resolution of the tryptic peptides of extracted proteins of the above three maize species was obtained in isoelectric BGE, 200 mM iminodiacetic acid, pH 2.26. Selected resolved tryptic peptides of proteins were characterised by effective electrophoretic mobilities and corrected (migration times normalised) peak areas. Some significant relative qualitative and quantitative differences in CZE-UV profiling of tryptic protein digests were found, which can be potentially used to differentiate transgenic Aristis Bt and non-transgenic Aristis varieties or two native non-transgenic varieties, Aristis and Coventry.