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Hyung Joon Cha - One of the best experts on this subject based on the ideXlab platform.
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enhanced production of dopa incorporated Mussel Adhesive protein using engineered translational machineries
Biotechnology and Bioengineering, 2020Co-Authors: Ye Seul Jeong, Hyung Joon Cha, Byeongseon Yang, Byungseop Yang, Mincheol Shin, Jihyoun Seong, Inchan KwonAbstract:Mussel Adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although in vivo residue-specific incorporation of 3,4-dihydroxyphenylalanine (Dopa) in tyrosine-auxotrophic Escherichia coli cells allows for production of Dopa-incorporated bioengineered MAPs (dMAPs), the low production yield hinders the practical application of dMAPs. This low production yield of dMAPs is due to low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, to enhance the production yield of dMAPs, we investigated the coexpression of Dopa-recognizing tyrosyl-tRNA synthetases (TyrRSs). To use the Dopa-specific Methanococcus jannaschii TyrRS (MjTyrRS-Dopa), we altered the anticodon of tyrosyl-tRNA amber suppressor into AUA (MjtRNATyr AUA ) to recognize a tyrosine codon (AUA). Co-overexpression of MjTyrRS-Dopa and MjtRNATyr AUA increased the production yield of Dopa-incorporated MAP foot protein type 3 (dfp-3) by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of dfp-3. Even with coexpression of Dopa-recognizing TyrRSs, dfp-3 has a high Dopa incorporation yield (over 90%) compared to ones prepared without TyrRS coexpression.
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Enhanced production of Dopa‐incorporated Mussel Adhesive protein using engineered translational machineries
Biotechnology and bioengineering, 2020Co-Authors: Ye Seul Jeong, Hyung Joon Cha, Byeongseon Yang, Byungseop Yang, Mincheol Shin, Jihyoun Seong, Inchan KwonAbstract:Mussel Adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although in vivo residue-specific incorporation of 3,4-dihydroxyphenylalanine (Dopa) in tyrosine-auxotrophic Escherichia coli cells allows for production of Dopa-incorporated bioengineered MAPs (dMAPs), the low production yield hinders the practical application of dMAPs. This low production yield of dMAPs is due to low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, to enhance the production yield of dMAPs, we investigated the coexpression of Dopa-recognizing tyrosyl-tRNA synthetases (TyrRSs). To use the Dopa-specific Methanococcus jannaschii TyrRS (MjTyrRS-Dopa), we altered the anticodon of tyrosyl-tRNA amber suppressor into AUA (MjtRNATyr AUA ) to recognize a tyrosine codon (AUA). Co-overexpression of MjTyrRS-Dopa and MjtRNATyr AUA increased the production yield of Dopa-incorporated MAP foot protein type 3 (dfp-3) by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of dfp-3. Even with coexpression of Dopa-recognizing TyrRSs, dfp-3 has a high Dopa incorporation yield (over 90%) compared to ones prepared without TyrRS coexpression.
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Mussel Adhesive protein-based whole cell array biosensor for detection of organophosphorus compounds
'Elsevier BV', 2018Co-Authors: Cs Kim, Bong-hyuk Choi, Jeong Hyun Seo, Lim G, Hyung Joon ChaAbstract:A whole cell array biosensor for the efficient detection of neurotoxic organophosphate compounds (OPs) was developed through the immobilization of recombinant Escherichia coli cells containing periplasmic-expressing organophosphorus hydrolase (OPH) onto the surface of a 96-well microplate using Mussel Adhesive protein (MAP) as a microbial cell-immobilizing linker. Both the paraoxon-hydrolyzing activity and fluorescence microscopy analyses demonstrated that the use of MAP in a whole cell biosensor increased the cell-immobilizing efficiency and enhanced the stability of immobilized cells compared to a simple physical adsorption-based whole cell system. Scanning electron microscopic analyses also showed that the E. coli cells were effectively immobilized on the MAP-coated surface without any pretreatment steps. The whole cell array biosensor system, prepared using optimal MAP coating (50 mu g/cm(2)) and cell loading (4 OD600), detected paraoxon levels as low as 5 mu M with high reproducibility, and its quantitative detection range was similar to 5-320 mu M. The MAP-based whole cell array biosensor showed a good long-term stability for 28 day with 80% retained activity and a reusability of up to 20 times. In addition, paraoxon in tap water was also successfully detected without a reduction in sensitivity. Our results indicate that the proposed MAP-based whole cell array system could be used as a potential platform for a stable and reusable whole cell biosensor. (C) 2012 Elsevier B.V. All rights reserved.X1202
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Facile Surface Functionalization with Glycosaminoglycans by Direct Coating with Mussel Adhesive Protein
'Mary Ann Liebert Inc', 2018Co-Authors: Bh Choi, Ds Hwang, Yoo Sung Choi, Hyung Joon ChaAbstract:The use of Mussel Adhesive proteins (MAPs) as a surface coating for cell adhesion has been suggested due to their unique properties of biocompatibility and effective adhesion on diverse inorganic and organic surfaces. The surface functionalization of scaffolds or implants using extracellular matrix (ECM) molecules is important for the enhancement of target cell behaviors such as proliferation and differentiation. In the present work, we suggest a new, simple surface functionalization platform based on the charge interactions between the positively charged MAP linker and negatively charged ECM molecules, such as glycosaminoglycans (GAGs). MAP was efficiently coated onto a titanium model surface using its adhesion ability. Then, several GAG molecules, including hyaluronic acid (HA), heparin sulfate (HS), chondroitin sulfate (CS), and dermatan sulfate (DS), were effectively immobilized on the MAP-coated surfaces by charge interactions. Using HA as a model GAG molecule, we found that the proliferation, spreading, and differentiation behaviors of mouse preosteoblast cells were all significantly improved on MAP/HA-layered titanium. In addition, we successfully constructed a multilayer film on a titanium surface with oppositely charged layer-by-layer coatings of MAP and HA. Collectively, our simple MAP-based surface functionalization strategy can be successfully used for the efficient surface immobilization of negatively charged ECM molecules in various tissue engineering and medical implantation applications.X1131
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Complex coacervates based on recombinant Mussel Adhesive proteins: their characterization and applications
Soft matter, 2017Co-Authors: Hyo Jeong Kim, Seonghye Lim, Byeongseon Yang, Tae Yoon Park, Hyung Joon ChaAbstract:Complex coacervates are a dense liquid phase of oppositely charged polyions formed by the associative separation of a mixture of polyions. Coacervates have been widely employed in many fields including the pharmaceutical, cosmetic, and food industries due to their intriguing interfacial and bulk material properties. More recently, attempts to develop an effective underwater Adhesive have been made using complex coacervates that are based on recombinant Mussel Adhesive proteins (MAPs) due to the water immiscibility of complex coacervates and the Adhesiveness of MAPs. MAP-based complex coacervates contribute to our understanding of the physical nature of complex coacervates and they provide a promising alternative to conventional invasive surgical repairs. Here, this review provides an overview of recombinant MAP-based complex coacervations, with an emphasis on their characterization and the uses of such materials for applications in the fields of biomedicine and tissue engineering.
Seonghye Lim - One of the best experts on this subject based on the ideXlab platform.
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RECOMBINANT Mussel Adhesive PROTEIN AS A GENE DELIVERY MATERIAL
'Wiley', 2019Co-Authors: Ds Hwang, Yoo Seong Choi, Kyoung Ro Kim, Seonghye Lim, Cha, Hyung JoonAbstract:Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant Mussel Adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine (TM) 2000. Our results indicate that this Mussel Adhesive protein may be used as a potential protein-based gene-transfer mediator.Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant Mussel Adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine (TM) 2000. Our results indicate that this Mussel Adhesive protein may be used as a potential protein-based gene-transfer mediator.X1
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Bulk Adhesive strength of recombinant hybrid Mussel Adhesive protein
'Informa UK Limited', 2018Co-Authors: Hj Cha, Ds Hwang, Seonghye Lim, James D. White, Cristina R. Matos-pérez, Jonathan J WilkerAbstract:Mussel Adhesive proteins (MAPs) have received increased attention as potential biomedical and environmental friendly Adhesives. However, practical application of MAPs has been severely limited by uneconomical extraction and unsuccessful genetic production. Developing new Adhesives requires access to large quantities of material and demonstrations of bulk mechanical properties. Previously, the authors designed fp-151, a fusion protein comprised of six MAP type 1 (fp-1) decapeptide repeats at each MAP type 5 (fp-5) terminus and successfully expressed it in Escherichia coli. This recombinant hybrid protein exhibited high-level expression, a simple purification and high biocompatibility as well as strong Adhesive ability on a micro-scale. In the present work, investigations on the bulk Adhesive properties of semi-purified (90% purity) fusion fp-151 were performed in air. The unmodified recombinant fp-151, as expressed, contains tyrosine residues and showed significant shear-Adhesive forces (0.33MPa). Adhesion strength increased (0.45MPa) after enzymatic oxidation of tyrosine residues to l-3,4-dihydroxyphenylalanine (DOPA) groups. Addition of cross-linkers such as iron(III), manganese(III) and periodate (IO4-) generally enhanced adhesion, although too much addition decreased adhesion. Among the three cross-linking reagents examined, the non-metallic oxidant periodate showed the highest shear-Adhesive forces (0.86MPa). In addition, it was found that Adhesive strengths could be increased by adding weights to the samples. The highest adhesion strength found was that of DOPA-containing fp-151 cross-linked with periodate and having weights applied to the samples (1.06MPa). Taken together, the first bulk-scale Adhesive force measurements are presented for an expressed recombinant hybrid Mussel Adhesive protein.X1393
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Complex coacervates based on recombinant Mussel Adhesive proteins: their characterization and applications
Soft matter, 2017Co-Authors: Hyo Jeong Kim, Seonghye Lim, Byeongseon Yang, Tae Yoon Park, Hyung Joon ChaAbstract:Complex coacervates are a dense liquid phase of oppositely charged polyions formed by the associative separation of a mixture of polyions. Coacervates have been widely employed in many fields including the pharmaceutical, cosmetic, and food industries due to their intriguing interfacial and bulk material properties. More recently, attempts to develop an effective underwater Adhesive have been made using complex coacervates that are based on recombinant Mussel Adhesive proteins (MAPs) due to the water immiscibility of complex coacervates and the Adhesiveness of MAPs. MAP-based complex coacervates contribute to our understanding of the physical nature of complex coacervates and they provide a promising alternative to conventional invasive surgical repairs. Here, this review provides an overview of recombinant MAP-based complex coacervations, with an emphasis on their characterization and the uses of such materials for applications in the fields of biomedicine and tissue engineering.
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Interfacial Tension of Complex Coacervated Mussel Adhesive Protein According to the Hofmeister Series
Langmuir : the ACS journal of surfaces and colloids, 2014Co-Authors: Seonghye Lim, Jeong Hyun Seo, Dustin Moon, Hyo Jeong Kim, In Seok Kang, Hyung Joon ChaAbstract:Complex coacervation is a liquid–liquid phase separation in a colloidal system of two oppositely charged polyelectrolytes or colloids. The interfacial tension of the coacervate phase is the key parameter for micelle formation and interactions with the encapsulating material. However, the relationship between interfacial tensions and various salt solutions is poorly understood in complex coacervation. In the present work, the complex coacervate dynamics of recombinant Mussel Adhesive protein (MAP) with hyaluronic acid (HA) were determined in the presence of Hofmeister series salt ions. Using measurements of absorbance, hydrodynamic diameter, capillary force, and receding contact angle in the bulk phase, the interfacial tensions of complex coacervated MAP/HA were determined to be 0.236, 0.256, and 0.287 mN/m in 250 mM NaHCOO, NaCl, and NaNO3 solutions, respectively. The sequences of interfacial tensions and contact angles of the complex coacervates in the presence of three sodium salts with different anions...
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in vivo post translational modifications of recombinant Mussel Adhesive protein in insect cells
Biotechnology Progress, 2011Co-Authors: Seonghye Lim, Yoo Seong Choi, Kyoung Ro Kim, Dae-kyum Kim, Daehee Hwang, Hyung Joon ChaAbstract:Mussel Adhesive proteins (MAPs) have been suggested as promising bioAdhesives for diverse application fields, including medical uses. Previously, we successfully constructed and produced a new type of functional recombinant MAP, fp-151, in a prokaryotic Escherichia coli expression system. Even though the E. coli-derived MAP showed several excellent features, such as high production yield and efficient purification, in vitro enzymatic modification is required to convert tyrosine residues to l-3,4-dihydroxyphenyl alanine (dopa) molecules for its Adhesive ability, due to the intrinsic inability of E. coli to undergo post-translational modification. In this work, we produced a soluble recombinant MAP in insect Sf9 cells, which are widely used as an effective and convenient eukaryotic expression system for eukaryotic foreign proteins. Importantly, we found that insect-derived MAP contained converted dopa residues by in vivo post-translational modification. In addition, insect-derived MAP also had other post-translational modifications including phosphorylation of serine and hydroxylation of proline that originally occurred in some natural MAPs. To our knowledge, this is the first report on in vivo post-translational modifications of MAP containing dopa and other modified amino acid residues.
Yoo Seong Choi - One of the best experts on this subject based on the ideXlab platform.
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RECOMBINANT Mussel Adhesive PROTEIN AS A GENE DELIVERY MATERIAL
'Wiley', 2019Co-Authors: Ds Hwang, Yoo Seong Choi, Kyoung Ro Kim, Seonghye Lim, Cha, Hyung JoonAbstract:Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant Mussel Adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine (TM) 2000. Our results indicate that this Mussel Adhesive protein may be used as a potential protein-based gene-transfer mediator.Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant Mussel Adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine (TM) 2000. Our results indicate that this Mussel Adhesive protein may be used as a potential protein-based gene-transfer mediator.X1
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Mussel Adhesive protein as an environmentally-friendly harmless wood furniture Adhesive
International Journal of Adhesion and Adhesives, 2016Co-Authors: Young Hoon Song, Yoo Seong Choi, Bong-hyuk ChoiAbstract:Abstract Recent Adhesive technologies have focused on the development of high-quality and eco-friendly Adhesives. Thus, there is a gradual shift from the currently used chemical-based Adhesives toward harmless Adhesives with improved quality and performance. Here, we evaluated the potential use of bacteria-produced recombinant Mussel Adhesive protein (MAP) as a harmless wood furniture Adhesive. We formulated a MAP wood Adhesive as an inclusion body type for economical preparation, and we confirmed its harmlessness through the non-detection of volatile organic compounds and heavy metals. The formulated MAP showed sufficiently strong bulk Adhesive strength for the dried gluing of wood adherends. We also found that the formulated MAP wood Adhesive exhibits robust adhesion in various environmental conditions, including open assembly times, incubation times, temperatures, and humidity levels. In summary, the developed recombinant MAP could be successfully used as a promising environmentally-friendly, harmless wood furniture Adhesive.
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A comparative study on the bulk Adhesive strength of the recombinant Mussel Adhesive protein fp-3
Biofouling, 2013Co-Authors: Byeongseon Yang, Yoo Seong Choi, Jeong Hyun Seo, Dong Gyun Kang, Hyung Joon ChaAbstract:Mussel Adhesive protein (MAP) type 3 (fp-3) is considered one of the key components for Mussel adhesion. However, its bulk Adhesive strength has not been characterized due to its availability in limited quantities. In the present work, a feasible production (∼47 mg l−1) of recombinant fp-3 was achieved, and its bulk Adhesive strength was measured for the first time; ∼0.57 MPa for the unmodified form and ∼0.94 and ∼2.28 MPa for the 3,4-dihydroxy-L-phenylalanine (DOPA)-modified form, having a 9.6% yield without and with oxidant treatment, respectively. Furthermore, values for the bulk Adhesive strength of several DOPA-modified recombinant MAPs were compared. The maximum Adhesive strength of DOPA-modified fp-3 after oxidant treatment was stronger than that of type 5 (fp-5), which has a 6.2% modification yield, and was comparable to that of hybrid types fp-131 and fp-151, which have similar yields (∼5%). The strong bulk Adhesive property of recombinant fp-3 demonstrates its potential use as a promising bioadh...
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Reinforced Multifunctionalized Nanofibrous Scaffolds Using Mussel Adhesive Proteins
Angewandte Chemie (International ed. in English), 2011Co-Authors: Bum Jin Kim, Yoo Seong Choi, Hyung Joon ChaAbstract:Nanofiber technology has been largely recognized because of its ability to fabricate multiple nanosized fibers that are structurally similar to collagen fibrils of the natural extracellular matrix (ECM). The large surface area of nanofibers is a major advantage of nanofibrous scaffolds because these are the sites at which cell–substrate interactions occur. Rendering the nanofiber surface to be biofunctional is critical for its successful application in tissue engineering. Synthetic and natural biomaterials have been widely used for the fabrication of nanofibrous scaffolds for tissue engineering applications. Blending synthetic and natural biomaterials has been used to combine the excellent biological activities of natural biomaterials with the high processability and desired mechanical strengths of synthetic polymers; however, this approach has had limited success for several pairs of synthetic and natural biomaterials. Several bioactive molecules, such as ECM proteins, ECM carbohydrates, ECM-derived peptides, and growth factors, have been introduced onto the surface of nanofibers to design biofunctional and biomimetic tissue engineering scaffolds. Generally, typical conjugation chemistry and physical adsorption procedures have been used to immobilize biomolecules after the completion of the surface activation steps, including plasma and wet-chemical treatments or surface graft polymerization. However, these strategies are limited in availability because of the multiple, complicated procedures that are required to complete them. Thus, development of a simple and effective surface functionalization strategy for nanofibers would be a promising tool for successful tissue engineering applications. In the present work, we propose to use Mussel Adhesive protein (MAP) as a natural biomaterial that serves as a blending partner for the preparation of sticky nanofibrous scaffolds. Using MAPs as a blending partner provides a facile, efficient, and multifunctionalizing platform for generating novel nanofibrous scaffolds. In fact, nanofibrous scaffolds based on MAPs having mechanical reinforcement were fabricated by a simple e lectrospinning process to develop a “general coating platform” for diverse bioactive molecules (Figure 1).
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in vivo post translational modifications of recombinant Mussel Adhesive protein in insect cells
Biotechnology Progress, 2011Co-Authors: Seonghye Lim, Yoo Seong Choi, Kyoung Ro Kim, Dae-kyum Kim, Daehee Hwang, Hyung Joon ChaAbstract:Mussel Adhesive proteins (MAPs) have been suggested as promising bioAdhesives for diverse application fields, including medical uses. Previously, we successfully constructed and produced a new type of functional recombinant MAP, fp-151, in a prokaryotic Escherichia coli expression system. Even though the E. coli-derived MAP showed several excellent features, such as high production yield and efficient purification, in vitro enzymatic modification is required to convert tyrosine residues to l-3,4-dihydroxyphenyl alanine (dopa) molecules for its Adhesive ability, due to the intrinsic inability of E. coli to undergo post-translational modification. In this work, we produced a soluble recombinant MAP in insect Sf9 cells, which are widely used as an effective and convenient eukaryotic expression system for eukaryotic foreign proteins. Importantly, we found that insect-derived MAP contained converted dopa residues by in vivo post-translational modification. In addition, insect-derived MAP also had other post-translational modifications including phosphorylation of serine and hydroxylation of proline that originally occurred in some natural MAPs. To our knowledge, this is the first report on in vivo post-translational modifications of MAP containing dopa and other modified amino acid residues.
Dong Soo Hwang - One of the best experts on this subject based on the ideXlab platform.
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cation π interaction in dopa deficient Mussel Adhesive protein mfp 1
Journal of Materials Chemistry B, 2015Co-Authors: Sangsik Kim, Ali Faghihnejad, Yongjin Lee, Yongseok Jho, Hongbo Zeng, Dong Soo HwangAbstract:Here we report the possible contribution of cation–π interaction to underwater adhesion of Mussels by using DOPA-deficient recombinant Mussel Adhesive proteins. Considering the instability of DOPA in an oxidative environment, the cation–π interaction in DOPA-deficient biopolymers provides a complementary cross-linking mechanism for the design of novel underwater Adhesives.
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promotion of osteoblast proliferation on complex coacervation based hyaluronic acid recombinant Mussel Adhesive protein coatings on titanium
Biomaterials, 2010Co-Authors: Dong Soo Hwang, Herbert J Waite, Matthew TirrellAbstract:Many biological polyelectrolytes are capable of undergoing a fluid–fluid phase separation known as complex coacervation. Coacervates were prepared using hyaluronic acid (HA) and a recombinant fusion protein consisting of Mussel Adhesive motifs and the RGD peptide (fp-151-RGD). The low interfacial energy of the coacervate was exploited to coat titanium (Ti), a metal widely used in implant materials. The coacervate effectively distributed both HA and fp-151-RGD over the Ti surfaces and enhanced osteoblast proliferation. Approximately half of total fp-151-RGD and HA in the solution transferred to the titanium surface within 2 h. Titanium coated with coacervates having high residual negative surface charge showed the highest cell proliferation of preosteoblast cells (MC-3T3) compared to the treatments tested. Indeed, MC-3T3 cells on complex coacervate coated titanium foils exhibited over 5 times greater cell proliferation than bare, HA coated or fp-151-RGD coated titanium.
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glycosylated hydroxytryptophan in a Mussel Adhesive protein from perna viridis
Journal of Biological Chemistry, 2009Co-Authors: Hua Zhao, Dong Soo Hwang, Jason Sagert, Herbert J WaiteAbstract:Abstract The 3,4-dihydroxyphenyl-l-alanine (Dopa)-containing proteins of Mussel byssus play a critical role in wet adhesion and have inspired versatile new synthetic strategies for Adhesives and coatings. Apparently, however, not all Mussel Adhesive proteins are beholden to Dopa chemistry. The cDNA-deduced sequence of Pvfp-1, a highly aromatic and redox active byssal coating protein in the green Mussel Perna viridis, suggests that Dopa may be replaced by a post-translational modification of tryptophan. The N-terminal tryptophan-rich domain of Pvfp-1 contains 42 decapeptide repeats with the consensus sequences ATPKPW1TAW2K and APPPAW1TAW2K. A small collagen domain (18 Gly-X-Y repeats) is also present. Tandem mass spectrometry of isolated tryptic decapeptides has detected both C2-hexosylated tryptophan (W1) and C2-hexosylated hydroxytryptophan (W2), the latter of which is redox active. The UV absorbance spectrum of W2 is consistent with 7-hydroxytryptophan, which represents an intriguing new theme for bioinspired opportunistic wet adhesion.
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Recombinant Mussel Adhesive protein as a gene delivery material.
Biotechnology and bioengineering, 2009Co-Authors: Dong Soo Hwang, Yoo Seong Choi, Kyoung Ro Kim, Seonghye Lim, Hyung Joon ChaAbstract:Efficient target gene delivery into eukaryotic cells is important for biotechnological research and gene therapy. Gene delivery based on proteins, including histones, has recently emerged as a powerful non-viral DNA transfer technique. Here, we investigated the potential use of a recombinant Mussel Adhesive protein, hybrid fp-151, as a gene delivery material, in view of its similar basic amino acid composition to histone proteins, and cost-effective and high-level production in Escherichia coli. After confirming DNA binding affinity, we transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign genes using hybrid fp-151 as the gene delivery carrier. Hybrid fp-151 displayed comparable transfection efficiency in both mammalian cell lines, compared to the widely used transfection agent, Lipofectamine 2000. Our results indicate that this Mussel Adhesive protein may be used as a potential protein-based gene-transfer mediator.
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Bulk Adhesive strength of recombinant hybrid Mussel Adhesive protein.
Biofouling, 2009Co-Authors: Hyung Joon Cha, Seonghye Lim, Dong Soo Hwang, James D. White, Cristina R. Matos-pérez, Jonathan J WilkerAbstract:Mussel Adhesive proteins (MAPs) have received increased attention as potential biomedical and environmental friendly Adhesives. However, practical application of MAPs has been severely limited by u...
Jonathan J Wilker - One of the best experts on this subject based on the ideXlab platform.
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Bulk Adhesive strength of recombinant hybrid Mussel Adhesive protein
'Informa UK Limited', 2018Co-Authors: Hj Cha, Ds Hwang, Seonghye Lim, James D. White, Cristina R. Matos-pérez, Jonathan J WilkerAbstract:Mussel Adhesive proteins (MAPs) have received increased attention as potential biomedical and environmental friendly Adhesives. However, practical application of MAPs has been severely limited by uneconomical extraction and unsuccessful genetic production. Developing new Adhesives requires access to large quantities of material and demonstrations of bulk mechanical properties. Previously, the authors designed fp-151, a fusion protein comprised of six MAP type 1 (fp-1) decapeptide repeats at each MAP type 5 (fp-5) terminus and successfully expressed it in Escherichia coli. This recombinant hybrid protein exhibited high-level expression, a simple purification and high biocompatibility as well as strong Adhesive ability on a micro-scale. In the present work, investigations on the bulk Adhesive properties of semi-purified (90% purity) fusion fp-151 were performed in air. The unmodified recombinant fp-151, as expressed, contains tyrosine residues and showed significant shear-Adhesive forces (0.33MPa). Adhesion strength increased (0.45MPa) after enzymatic oxidation of tyrosine residues to l-3,4-dihydroxyphenylalanine (DOPA) groups. Addition of cross-linkers such as iron(III), manganese(III) and periodate (IO4-) generally enhanced adhesion, although too much addition decreased adhesion. Among the three cross-linking reagents examined, the non-metallic oxidant periodate showed the highest shear-Adhesive forces (0.86MPa). In addition, it was found that Adhesive strengths could be increased by adding weights to the samples. The highest adhesion strength found was that of DOPA-containing fp-151 cross-linked with periodate and having weights applied to the samples (1.06MPa). Taken together, the first bulk-scale Adhesive force measurements are presented for an expressed recombinant hybrid Mussel Adhesive protein.X1393
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high strength underwater bonding with polymer mimics of Mussel Adhesive proteins
ACS Applied Materials & Interfaces, 2017Co-Authors: Michael A North, Chelsey A Del Grosso, Jonathan J WilkerAbstract:When it comes to underwater adhesion, shellfish are the true experts. Mussels, barnacles, and oysters attach to rocks with apparent ease. Yet our man-made glues often fail when trying to stick in wet environments. Results described herein focus on a copolymer mimic of Mussel Adhesive proteins, poly(catechol-styrene). Underwater bonding was examined as a function of parameters including polymer molecular weight and composition. In doing so, several surprising results emerged. Poly(catechol-styrene) may be the strongest underwater Adhesive found to date. Bonding even exceeded that of the reference biological system, live Mussels. Adhesion was also found to be stronger under salt water than deionized water. Such unexpected findings may contradict an earlier proposal in which charged amino acids were suggested to be key for Mussel Adhesive function. Taken together, these discoveries are helping us to both understand biological adhesion as well as develop new materials with properties not accessed previously.
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Combining biomimetic principles from the lotus leaf and Mussel Adhesive: polystyrene films with superhydrophobic and Adhesive layers
RSC Advances, 2013Co-Authors: Ana I. Neto, Courtney L Jenkins, Heather J Meredith, Jonathan J Wilker, João Filipe ManoAbstract:Lotus leaves are well known for their extremely water repellent surfaces. Marine Mussels are also a popular research topic when considering biological Adhesives. Both organisms have inspired the development of several biomimetic materials. Herein we describe a two-sided film made almost entirely from polystyrene onto which the properties of both lotus leaves and Mussel Adhesive are incorporated. On one side of the film, imparting micrometer and nanometer scale hierarchical roughness yields superhydrophobicity and water repellency, which facilitates rapid fluid flow. The other side of the film is modified with a copolymer mimic of 3,4-dihydroxyphenylalanine (DOPA)-containing Mussel Adhesive proteins. This copolymer incorporates 3,4-dihydroxystyrene, to represent DOPA, randomly into a polystyrene host polymer. The flexibility of the polystyrene backing film enabled rolling of the assembly into a tubular shape. Inside the polystyrene tube was the superhydrophobic lotus mimic. The Mussel Adhesive mimic, on the outer layer, was used to glue the tube to itself, thus maintaining the tubular shape. The film was also successfully glued to a variety of flat substrates. These two-dimensional and three-dimensional assemblies can be used to direct and localize the flow of fluids, with partitioning between superhydrophobic and relatively hydrophilic regions. Such assemblies may facilitate the design of liquid transport for industrial and biomedical devices.
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Bulk Adhesive strength of recombinant hybrid Mussel Adhesive protein.
Biofouling, 2009Co-Authors: Hyung Joon Cha, Seonghye Lim, Dong Soo Hwang, James D. White, Cristina R. Matos-pérez, Jonathan J WilkerAbstract:Mussel Adhesive proteins (MAPs) have received increased attention as potential biomedical and environmental friendly Adhesives. However, practical application of MAPs has been severely limited by u...
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Thin films of polymer mimics of cross-linking Mussel Adhesive proteins deposited by matrix assisted pulsed laser evaporation
Applied Surface Science, 2009Co-Authors: R. Cristescu, Anand Doraiswamy, Jonathan J Wilker, Roger J. Narayan, Shane J. Stafslien, Ion N. Mihailescu, I Stamatin, Glenn Westwood, Bret J. Chisholm, Douglas B. ChriseyAbstract:Mussels secrete specialized Adhesives known as Mussel Adhesive proteins, which allow attachment of the organisms to underwater marine environments. Obtaining large quantities of naturally derived Mussel Adhesive proteins Adhesives has proven to date rather problematic, thus, synthetic analogs of Mussel Adhesive proteins have recently been developed. We report deposition of 1:100 and 1:1000 poly[(3,4-dihydroxystyrene)-co-styrene)] Mussel Adhesive protein analogs by matrix assisted pulsed laser evaporation (MAPLE) using an ArF* excimer laser source. The deposited films have been evaluated for their antifouling behavior. The MAPLE-deposited synthetic Mussel Adhesive protein analog thin films are homogenous and Adhesive, making the use of these materials in thin film form a viable option.
