A high adhesion cationic peptide

Abstract

The application discloses a high-adhesion cationic peptide. The amino acid sequence of the high-adhesion cationic peptide is K-Dopa-K-Dopa-K-Dopa-G-G-K-Y-G-G-K-Dopa-K-Dopa-K-Dopa-G-G-K-Y-G-G-K-Dopa-K-Dopa-K-Dopa. The high-adhesion cationic peptide has high adhesion strength, and can effectively bond various materials. The high-adhesion cationic peptide has the characteristics of high biocompatibility, good safety and strong adhesion, and can be applied to the fields of biological manufacturing and medicine.

Description

Technical Field

[0001] The present invention belongs to the field of biomimetic adhesion technology, and particularly relates to a polypeptide with high adhesion in a low pH environment and applications thereof. Background Art

[0002] Marine mussels, known for their ability to firmly adhere to various substrates through their secreted byssal protein (Mfp), have garnered widespread attention as adhesive materials for peptides inspired by Mfp. Studies have shown that Dopa (Dopa) in Mfp can attach to various substrate surfaces through adhesion mechanisms mediated by bidentate hydrogen bonds, metal-catechol coordination bonds, π-π / π-cation interactions, oxidative crosslinking, and electrostatic forces. Furthermore, lysine (Lys) in Mfp can expel hydrated cations from the substrate surface, promoting the binding of Dopa's catechol to the underlying substrate. Furthermore, literature review revealed that adjacent Lys residues in Mfp promote Dopa's underwater adhesion. Furthermore, incorporating additional Lys-Dopa adhesive sequences into the peptide structure did not significantly improve adhesion. For the Lys-Dopa repeat unit, (Lys-Dopa)3 reaches the upper limit of adhesion. Furthermore, increasing the number of tandem repeats does not significantly change adhesion, but does increase adhesion energy.

[0003] Studies have shown that in alkaline environments and under the influence of trace oxidants, Dopa readily undergoes two-electron oxidation to form dopaquinone, which reduces the number of Dopa residues available for binding and, in turn, the adhesion of Mfp. Therefore, by analyzing the sequence characteristics of Mfp, (Lys-Dopa)3 was identified as the primary adhesion sequence, linked with a flexible Gly and a protective Lys. By adjusting the (Lys-Dopa)3, Gly, and Lys residues in the peptide, a low-pH-adaptable, highly adhesive peptide was designed and screened, and its adhesion properties were analyzed. Summary of the Invention

[0004] The object of the present invention is to provide a polypeptide with high adhesion in a low pH environment and its application.

[0005] A highly adhesive cationic peptide, characterized in that its amino acid sequence is K-Dopa-K-Dopa-K-Dopa-GGKYGGK-Dopa-K-Dopa-K-Dopa-GGKYGGK-Dopa-K-Dopa-K-Dopa, wherein Dopa is L-3,4-dihydroxyphenylalanine.

[0006] The polypeptide has an adhesion pressure greater than 85 KPa and a microscopic adhesion force greater than 25 nN in an environment of pH 1-5.

[0007] Application of the highly adhesive cationic peptide in the preparation of highly adhesive biomaterials.

[0008] The application of the highly adhesive cationic peptide in the preparation of highly adhesive medical materials.

[0009] The beneficial effects of the present invention are as follows: The highly adhesive cationic peptides of the present invention have high adhesion and adsorption capacity, and can adhere to the surfaces of various organic and inorganic substances. The highly adhesive peptides of the present invention with these characteristics can be applied in fields such as biomanufacturing and medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 Adhesion pressure analysis of highly adhesive cationic peptides at different pH values.

[0011] Figure 2 Adhesion pressure analysis of high-adhesion cationic peptides with different curing times.

[0012] Figure 3 Microscopic adhesion analysis of highly adhesive cationic peptides. DETAILED DESCRIPTION

[0013] To facilitate understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0014] Example 1: Adhesion Pressure Analysis of High Adhesion Peptides at Different pH

[0015] Experimental steps:

[0016] 1. A highly adhesive cationic peptide sample (Nanjing Peptide Valley Biotechnology Co., Ltd., CAS: TG-LG-14188) was prepared into a peptide solution with a mass concentration of 5% using PBS buffer at pH 1, 3, 5, and 7, respectively.

[0017] 2. Add 1.4 μl of peptide solution to the surface of a 1 cm × 1.5 cm glass slide, quickly cover it with another glass slide of the same size and fix it with a clamp, and solidify it for 30 minutes.

[0018] 3. The shear pressure at different pH values ​​was measured using a tensile strength testing machine.

[0019] Experimental results:

[0020] Highly adhesive cationic peptides have good macroscopic adhesion in low pH environments, and the shear pressure does not change significantly. The adhesion is significantly reduced in a neutral environment. Figure 1 )

[0021] Example 2: Adhesion Pressure Analysis of Highly Adhesive Cationic Peptides at Different Curing Times

[0022] Experimental steps:

[0023] 1. Prepare a 5% mass concentration peptide solution of a highly adhesive cationic peptide and a control peptide sample (Nanjing Peptide Valley Biotechnology Co., Ltd., CAS: TG-LG-13650) with ultrapure water.

[0024] 2. Add 1.4 μl of peptide solution to the surface of a 1 cm × 1.5 cm glass slide, quickly cover it with another glass slide of the same size and fix it with a clamp, and cure it for 0.5 h and 2 h respectively.

[0025] 3. Use tensile strength testing machine to test the shear force of different curing times.

[0026] 4. Import the data into GraphPad Prism 8 for shear pressure analysis.

[0027] Experimental results:

[0028] The adhesion pressure of the highly adhesive cationic peptide increases with the extension of the curing time, and the adhesion pressure is greater than that of the control peptide. Figure 2 )

[0029] Example 3: Analysis of Microscopic Adhesion Force of Highly Adhesive Cationic Peptides

[0030] Experimental steps:

[0031] 1. Prepare a sample of the highly adhesive cationic peptide with distilled water to a concentration of 0.5%.

[0032] 2. Surface adhesion tests were conducted in air at a room temperature of 25°C and a relative humidity of 28%. The experimental instrument was a German Bruker FASTSCANBIO atomic force microscope. A FASTSCAN-C probe with a triangular Si3N4 tip, a radius of curvature of 5 nm, and an elastic modulus of 0.8 N / m was used. The probe's pressing force on the sample was set to 40 nN.

[0033] 3. Import the data into GraphPad Prism 8 and select points with similar microscopic adhesion for analysis.

[0034] Experimental results:

[0035] The microscopic average adhesion force of the low pH-adapted high-adhesion peptide can reach 27.75nN, which is higher than most adhesive materials reported in the literature. Figure 3 )

[0036] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A highly adhesive cationic peptide, characterized in that Its amino acid sequence is K-Dopa-K-Dopa-K-Dopa-GGKYGGK-Dopa-K-Dopa-K-Dopa-GGKYGGK-Dopa-K-Dopa-K-Dopa, where Dopa is L-3,4-dihydroxyphenylalanine. Under a pH environment of 1-5, the adhesion pressure of the polypeptide is higher than 85 kPa, and the microscopic adhesion force is higher than 25 nN.

2. Use of the highly adhesive cationic peptide according to claim 1 in the preparation of highly adhesive biomaterials.

3. Use of the highly adhesive cationic peptide according to claim 1 in the preparation of highly adhesive pharmaceutical materials.

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