Abstract
An anticancer bioactive peptide (ACBP), goat peroxiredoxin-5 (gPRDX5), was identified from goat-spleen extract after immunizing the goat with gastric cancer-cell lysate. Its amino acid sequence was determined by employing 2D nano-LC-ESI-LTQ-Orbitrap MS/MS combined with Mascot database search in the goat subset of the Uniprot database. The recombinant gPRDX5 protein was acquired by heterogeneous expression in Escherichia coli. Subsequently, the anti-cancer bioactivity of the peptide was measured by several kinds of tumor cells. The results indicated that the gPRDX5 was a good anti-cancer candidate, especially for killing B16 cells. However, the peptide was found to be unstable without modification with pharmaceutical excipients, which would be a hurdle for future medicinal application. In order to overcome this problem and find an effective way to evaluate the gPRDX5, nanoparticle formation, which has been widely used in drug delivery because of its steadiness in application, less side-effects and enhancement of drug accumulation in target issues, was used here to address the issues. In this work, the gPRDX5 was dispersed into nanoparticles before delivered to B16 cells. By the nanotechnological method, the gPRDX5 was stabilized by a fast and accurate procedure, which suggests a promising way for screening the peptide for further possible medicinal applications.
Highlights
The Nano-flow Liquid Chromatography in tandem with Orbitrap Mass Spectrum offers a good way for authentication purposes[14,15]
The goat peroxiredoxin-5 (gPRDX5) has attracted scientific interest in recent years since it was first identified by Su et al[10]
The gPRDX5 was first analyzed by Basic Local Alignment Search Tool (BLAST), http://blast.ncbi.nlm.nih.gov/Blast.cgi, and the results showed that its homogeneity to the peroxiredoxins V of human being was up to 89%
Summary
The Nano-flow Liquid Chromatography in tandem with Orbitrap Mass Spectrum offers a good way for authentication purposes[14,15]. It is significant to identify the amino acid sequence of gPRDX5 and enrich the protein by heterogeneous expression for further investigations Because of their poor stability, high variability, short half-lives and high molecular weight, the proteins/peptides are more sensitive to the change of enzymatic environment and pH than conventional drugs, and it is difficult for them to penetrate the intestinal mucosa by gastrointestinal administration. All these characteristics contribute to their limited medicinal application currently. The screening of anticancer bioactive proteins/peptides in combination with nanotechnology promises an effective method for medicinal purposes
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