Control of guanine-rich DNA secondary structures depending on the protease activity using a designed PNA peptide

“…These observations were verified by five macro-observation techniques, electrophoresis, DLS, zeta potential, CD, NMR, and gel filtration. The binding of the calmyc peptide and MYC had been assayed using electrophoresis in our previous paper [ 52 ]. Briefly, we checked that calmyc could bind to MYC, resulting in the cationic PNA peptide causing the bands to migrate with a higher range of apparent molecular weights than expected.…”

Section: Resultsmentioning

confidence: 99%

“…We previously constructed a regulation system for the DNA secondary structure formation of G-rich sequences, using a designed peptide nucleic acid (PNA) [ 45 , 46 , 47 , 48 , 49 , 50 , 51 ], whose functionality could be switched through protease activity [ 52 ] ( Figure 1 and Figure S1 ). Furthermore, this study implied that it was theoretically possible that G-wire structures constructed using this peptide could be changed to other nanostructures, and that the G-wire structure could be regenerated by the protease ( Figure 1 c).…”

Section: Introductionmentioning

confidence: 99%

DNA G-Wire Formation Using an Artificial Peptide is Controlled by Protease Activity

et al. 2017

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The development of a switching system for guanine nanowire (G-wire) formation by external signals is important for nanobiotechnological applications. Here, we demonstrate a DNA nanostructural switch (G-wire <--> particles) using a designed peptide and a protease. The peptide consists of a PNA sequence for inducing DNA to form DNA–PNA hybrid G-quadruplex structures, and a protease substrate sequence acting as a switching module that is dependent on the activity of a particular protease. Micro-scale analyses via TEM and AFM showed that G-rich DNA alone forms G-wires in the presence of Ca2+, and that the peptide disrupted this formation, resulting in the formation of particles. The addition of the protease and digestion of the peptide regenerated the G-wires. Macro-scale analyses by DLS, zeta potential, CD, and gel filtration were in agreement with the microscopic observations. These results imply that the secondary structure change (DNA G-quadruplex <--> DNA/PNA hybrid structure) induces a change in the well-formed nanostructure (G-wire <--> particles). Our findings demonstrate a control system for forming DNA G-wire structures dependent on protease activity using designed peptides. Such systems hold promise for regulating the formation of nanowire for various applications, including electronic circuits for use in nanobiotechnologies.

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“…[10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] (2) Peptides can be conjugated with articial amino acids and functional molecules. 19,[26][27][28] (3) Peptides are easier to design and handle than proteins. However, clear guidelines for the relationships between peptide sequences and inorganic precipitation ability have not been established.…”

Section: Introductionmentioning

confidence: 99%