title: 'Click' chemistry synthesis and capillary electrophoresis study of 1,4-linked 1,2,3-triazole AZT-systemin conjugate.
abstract: The Cu(I) catalyzed Huisgen 1,3-dipolar azide-alkyne cycloaddition (CuAAC) was applied for a nucleoside-peptide bioconjugation. Systemin (Sys), an 18-aa plant signaling peptide naturally produced in response to wounding or pathogen attack, was chemically synthesized as its N-propynoic acid functionalized analog (Prp-Sys) using the SPPS. Next, CuAAC was applied to conjugate Prp-Sys with 3'-azido-2',3'-dideoxythymidine (AZT), a model cargo molecule. 1,4-Linked 1,2,3-triazole AZT-Sys conjugate was designed to characterize the spreading properties and ability to translocate of cargo molecules of systemin. CuAAC allowed the synthesis of the conjugate in a chemoselective and regioselective manner, with high purity and yield. The presence of Cu(I) ions generated in situ drove the CuAAC reaction to completion within a few minutes without any by-products. Under typical separation conditions of phosphate 'buffer' at low pH and uncoated fused bare-silica capillary, an increasing peak intensity assigned to triazole-linked AZT-Sys conjugate was observed using capillary electrophoresis (CE) during CuAAC. CE analysis showed that systemin peptides are stable in tomato leaf extract for up to a few hours. CE-ESI-MS revealed that the native Sys and its conjugate with AZT are translocated through the tomato stem and can be directly detected in stem exudates. The results show potential application of systemin as a transporter of low molecular weight cargo molecules in tomato plant and of CE method to characterize a behavior of plant peptides and its analogs.
authors.affiliation: Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, Gdańsk, 80-308, Poland; Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Dobkowski
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Szychowska
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Pieszko
authors.forename: Anna
authors.initials: A
authors.lastname: Miszka
authors.forename: Monika
authors.initials: M
authors.lastname: Wojciechowska
authors.forename: Magdalena
authors.initials: M
authors.lastname: Alenowicz
authors.forename: Jarosław
authors.initials: J
authors.lastname: Ruczyński
authors.forename: Piotr
authors.initials: P
authors.lastname: Rekowski
authors.forename: Lech
authors.initials: L
authors.lastname: Celewicz
authors.forename: Jan
authors.initials: J
authors.lastname: Barciszewski
authors.forename: Piotr
authors.initials: P
authors.lastname: Mucha
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry*; D057930:Click Chemistry / Q000379:methods*; D004337:Drug Carriers / Q000138:chemical synthesis* / Q000737:chemistry; D019075:Electrophoresis, Capillary; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D060327:Solid-Phase Synthesis Techniques; D015215:Zidovudine / Q000737:chemistry*
chemical_list: D019380:Anti-HIV Agents; D004337:Drug Carriers; D010455:Peptides; C073704:systemin; D015215:Zidovudine
keywords: AZT; capillary electrophoresis; click chemistry; peptide-nucleoside bioconjugation; systemin