Applied Chemistry for Engineering, Vol.31, No.1, 67-72, February, 2020
Enzymatic Conjugation of RGD Peptides on the Surface of Fibroin Microspheres
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Biomaterials are frequently functionalized with Arg-Gly-Asp (RGD) peptides to provide cell adhesion sites. In this study, RGD peptides were enzymatically coupled on to the surface of fibroin microspheres. Papain exhibited a strong preference for dansyl phenylalanine for the peptide formation with fibroin microspheres. Thus, RGD1 peptide was designed to carry cysteine to both sides of the sequence, glycine as a spacer and two residues of phenylalanine at the C-terminal (CRGDCGFF). The enzymatic modification facilitated by an increasing amount of substrate and by the presence of organic solvent, dimethylsulfoxide at 25% (v/v). Microspheres coupled with RGD1, showed a significantly different precipitation property and an increased apparent volume, possibly due to the steric hindrance of RGD peptides on the surface. Transmission electron microscopy also confirmed the presence of cysteine residues in RGD1 coupled on the surface of microspheres stained with gold nanoparticles. RGD1-microspheres significantly facilitated the growth of murine fibroblast 3T3 cells even under non-adhesion culture conditions.
- Bellis S, Biomaterials, 32, 4205 (2011)
- Ruoslahti E, Pierschbacher MD, Science, 238, 491 (1987)
- Wang F, Li YY, Shen YQ, Wang AM, Wang SL, Xie T, Int. J. Mol. Sci., 14(7), 13447 (2013)
- Hersel U, Dahmen C, Kessler H, Biomaterials, 24(24), 4385 (2003)
- Carpino LA, Shroff H, Triolo SA, Mansour EME, Wenschuh H, Albericio F, Tetrahedron Lett., 34, 7829 (1993)
- Hern DL, Hubbell JA, J. Biomed. Mater. Res., 39, 266 (1998)
- Lateef SS, Boateng S, Hartman TJ, Crot CA, Russell B, Hanley L, Biomaterials, 23, 3159 (2002)
- Vepari C, Kaplan DJ, Prog. Polym. Sci, 32(8-9), 991 (2007)
- Kim EJ, Lee SY, Hur W, J. Appl. Polym. Sci., 121(6), 3460 (2011)
- Ulijn RV, Baragana B, Halling PJ, Flitsch SL, J. Am. Chem. Soc., 124(37), 10988 (2002)
- Schuster M, Munoz B, Yuan W, Wong CH, Tetrahedron Lett., 34(8), 1247 (1993)
- Riener CK, Kada G, Gruber HJ, Anal. Bioanal. Chem., 373(4-5), 266 (2002)
- Walker JM, Basic Protein and Peptide Protocols, 321-328, Humana Press (1994).
- Zhou CZ, Confalonieri F, Medina N, Zivanovic Y, Esnault C, Yang T, Janin J, Duguet M, Perasso R, Li ZG, Nucleic Acids Res., 28, 2413 (2000)
- Morihara K, Oka T, J. Biochem., 89(2), 85 (1981)
- Anderson G, Luisi PL, Helv. Chim. Acta, 62, 488 (1979)
- Lecaille F, Serveau C, Gauthier F, Lalmanach G, Febs Lett., 445(2-3), 311 (1999)
- Leary R, Larsen D, Watanabe H, Shaw E, Biochemistry, 16(26), 5857 (1977)
- Mitin YV, Braun K, Kuhl P, Biotechnol. Bioeng., 54(3), 287 (1997)
- O’Neill CH, Riddle PN, Jordan PW, Cell, 16(4), 909 (1979)
- Bellis SL, Biomaterials, 32, 4205 (2011)