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Research Article

SRP. 2020; 11(9): 503-509


Use of Natural Peptides with a Cysteine Knot of Arthropods as a Carrier of a Peptide Tropic to Glutamate Carboxypeptidase II

Elena Iurova, Ivan Beloblov, Evgenii Beloborodov, Eugenia Rastorgueva, Evgenia Pogodina, Elizaveta Tazintseva, Yuri Saenko, Aleksandr Fomin.


Abstract

The aim of the investigation. Synthesis of a peptide with a cysteine knot, affinity to glutamate carboxypeptidase II, evaluation of its binding to glutamate carboxypeptidase II on the surface of LNCaP prostate cancer cells. Comparison of peptide binding to a cysteine knot, affinity to glutamate carboxypeptidase II with the parent peptide, affinity to glutamate carboxypeptidase II.
Materials and Methods. Peptide with sequense GTIQPYPFSWGY, affinity to glutamate carboxypeptidase II, was inserted into the peptide with the cysteine knot U5-Sth1a (isolated from the spider venom of the family Scytodes thoracica). The peptides were synthesized using a peptide synthesizer (ResPepSL, Intavis), the synthesis result was monitored by high performance liquid chromatography (NGC Quest, Bio-Rad) and mass spectrometry (MALDI-TOF MS, Bruker Daltonics). As an object of study, we used prostate cancer cell cultures LNCaP (synthesizes glutamate carboxypeptidase II on the surface), PC3 and DU145 (they do not synthesize glutamate carboxypeptidase II). In the course of the study, the initial tropic glutamate carboxypeptidase II peptide and hybrid peptide were labeled with 6-FAM NHS fluorescent dye, then added to the cell culture medium, then fluorescence was evaluated and compared.
Results. Greater binding of the peptide to the cysteine knot with glutamate carboxypeptidase II on the surface of LNCaP culture cells was noted, compared with the original peptide, affinity to glutamate carboxypeptidase II. An increase in the binding of the hybrid peptide to glutamate carboxypeptidase II with time, in the range from one hour to one day, was also noted.

Key words: Peptides, prostate cancer, targeted medicine, glutamate carboxypeptidase II, PSMA






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