NOVEL MATERIAL: BIOCOMPATIBLE GLUE FOR USE IN BIOLOGY AND MEDICINE—RECOMBINANT MUSSELADHESIVE PROTEINS
Main Article Content
Authors
A.S. Aхambayeva
National Center for Biotechnology, Korgalzhyn hwy, 13/5, Astana, 010000, Kazakhstan
Zh.S. Shagyrova
National Center for Biotechnology, Korgalzhyn hwy, 13/5, Astana, 010000, Kazakhstan
T.S. Nurgozhin
Nazarbayev University, Kabanbai Batyr avenue, 53, Astana, 010000, Kazakhstan
E. Zhienbay
Nazarbayev University, Kabanbai Batyr avenue, 53, Astana, 010000, Kazakhstan
A.V. Shustov
National Center for Biotechnology, Korgalzhyn hwy, 13/5, Astana, 010000, Kazakhstan
Abstract
Mussel adhesive proteins (MFPs) help mussels to attach to various surfaces. Natural MFPs present in the adhesive discs of the byssal threads of the mussels exhibit a number of good adhesiveproperties thatcan be used in medicine, orthodontics, and cell and tissue engineering. MFPs can be used to glue a variety of materials, natural and artificial. For use in medicine, it is important that the MFPs are waterproof adhesives, effectively bonding surfaces submerged in water. Natural MFPs are already being used as adhesives, but their utilization is limited because of high costs.
Recombinant MFPs are an attractive alternative because they can be produced in largequantities. For the recombinant MFPs produced by Escherichia coli to possess adhesive properties, post-translational modification of tyrosine residues to 3,4-dihydroxyphenylalanine (DOPA) is required.
This paper describes an expression system in which the recombinant adhesive protein Fp-131 is produced, and the newly synthesized polypeptide undergoes modification of the tyrosine residues to DOPA. Hydroxylation of tyrosine occurredin vivo in the bacterial cells because of the activity of tyrosinase, which is coexpressed with Fp-131. Coexpression of Fp-131 and tyrosinase was achieved through the expression of the proteins from two plasmids with different origins of replication and resistance markers.
Fp-131 was purified using metal affinity chromatography under denaturing conditions. After dialysis and freeze-drying, a product was obtained with an yield of 25 mg from 1 L of the induced culture. Presence of DOPA was demonstrated in Fp-131 by using a colour reaction with NBT. The adhesion strength of Fp-131 was measured using the lap-shear test. For this test, Fp-131 was used to glue two flat adherends, and a shear load was attached to the glued joint. The adhesive strength was 1.1 MPa, which is comparable to that of natural MFPs.
Keywords
adhesive protein, mussels, tyrosinase, DOPA, adhesion strength, biocompatible glue
Article Details
References
Stewart R.J., Ransom T.C., Hlady V.J. Natural Underwater Adhesives.PolymSci B Polym Phys, 2011,vol. 11, no. 9, pp. 757-771.
Lee B.P., Messersmith P.B., Israelachvili J.N., Waite J.H. Mussel-Inspired Adhesives and Coatings. Annu Rev Mater Res, 2011,vol. 41, no. 1,pp. 99-132.
Silverman H.G., Roberto F.F. Understanding marine mussel adhesion.Mar Biotechnol (NY), 2007,vol. 6, no. 9, pp. 661-681.
Jonker J.L., Abram F., Pires E., Varela Coelho A., Grunwald I. Adhesive proteins of stalked and acorn barnacles display homology with low sequence similarities.PLoS One, 2014,vol. 10, no. 8, 9,pp. 108-122.
Stewart R.J., Wang C.S., Song I.T., Jones J.P. The role of coacervation and phase transitions in the sandcastle worm adhesive system.Adv Colloid Interface Sci, 2016,vol. S0001, pp. 8686-8699.
Hatano T., Nagashima T. The Secretion Process of Liquid Silk with Nanopillar Structures from Stenopsychemarmorata(Trichoptera: Stenopsychidae). Sci. Rep., 2015, vol. 5, no.9237, pp. 1010-1038.
Lin Q., Gourdon D., Sun C., et al. Adhesion mechanisms of the mussel foot proteins mfp-1 and mfp-3.Proc. Natl. Acad. Sci. USA, 2007,vol. 10, pp. 3782-3786.
Clancy S.K., Sodano A., Cunningham D.J., et al. Marine Bioinspired Underwater Contact Adhesion. Biomacromolecules, 2016, vol. 5, no.17, pp. 1869-1874.
Li A., Mu Y., Jiang W., Wan X. A mussel-inspired adhesive with stronger bonding strength under underwater conditions than under dry conditions.ChemCommun (Camb), 2015, vol. 44, no. 51, pp. 9117-9120.
Lu Q., Danner E., Waite J.H., Israelachvili J.N., Zeng H., Hwang D.S. Adhesion of mussel foot proteins to different substrate surfaces. JR Soc Interface, 2013, vol. 79, no. 10, pp. 2641-2659.
Yu J., Kan Y., Rapp M., et al. Adaptive hydrophobic and hydrophilic interactions of mussel foot proteins with organic thin films. Proc. Natl. Acad. Sci. USA, 2013,vol. 39, no. 110, pp. 15680-15685.
Yu J., Wei W., Menyo M.S., et al. Adhesion of mussel foot protein-3 to TiO2 surfaces: the effect of pH.Biomacromolecules, 2013,vol. 4, no.14, pp. 1072-1077.
Haemers S., Koper G.J., Frens G. Effect of oxidation rate on cross-linking of mussel adhesive proteins. Biomacromolecules, 2003,vol. 3, no. 4, pp. 632-640.
Monahan J., Wilker J.J. Cross-linking the protein precursor of marine mussel adhesives: bulk measurements and reagents for curing.Langmuir, 2004,vol. 9, no. 20, pp. 3724-3729.
Krishnan V., Clark R., Chekmareva M., et al. In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose.J Vis Exp, 2015,vol. 105, pp. 52-72.
Hwang D.S., Yoo H.J., Jun J.H., et al. Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli.Appl Environ Microbiol, 2004,vol. 6, no. 70, pp. 3352-3359.
Hwang D.S., Gim Y., Cha H.J. Expression of functional recombinant mussel adhesive protein type 3A in Escherichia coli. BiotechnolProg, 2005,vol. 2, no. 21, pp. 965-970.
Briceño A., Muñoz P., Brito P., et al. Aminochrome Toxicity is Mediated by Inhibition of Microtubules Polymerization Through the Formation of Adducts with Tubulin. Neurotox Res, 2016,vol. 3, no. 29, pp. 381-393.
Yang B., Kang D.G., Seo J.H., et al. A comparative study on the bulk adhesive strength of the recombinant mussel adhesive protein fp-3.Biofouling, 2013, vol. 5, no. 29, pp. 483-490.
Choi Y.S., Yang Y.J., Yang B., Cha H.J. In vivo modification of tyrosine residues in recombinant mussel adhesive protein by tyrosinase co-expression in Escherichia coli.Microb Cell Fact, 2012, vol. 11, pp. 139-156.
Ren Q., Henes B., Fairhead M., Thöny-Meyer L. High level production of tyrosinase in recombinant Escherichia coli. BMC Biotechnol, 2013, vol. 13, pp. 18-36.
Chang T.S. An updated review of tyrosinase inhibitors.Int J MolSci, 2009,vol. 10, no.6, pp. 2440-2475.
Rodgers K.J., Hume P.M., Dunlop R.A., Dean R.T. Biosynthesis and turnover of DOPA-containing proteins by human cells.Free RadicBiol Med, 2004,vol. 37, no. 11, pp. 1756-1764.
Nicklisch S.C., Waite J.H. Mini-review: the role of redox in Dopa-mediated marine adhesion. Biofouling, 2012, vol. 28, no. 8, pp. 865-877.
Zhou J., Defante A.P., Lin F., et al. Adhesion properties of catechol-based biodegradable amino acid-based poly(ester urea) copolymers inspired from mussel proteins. Biomacromolecules, 2015,vol. 16, no. 1, pp. 266-274.