• Title/Summary/Keyword: Protein Crosslinking

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Anti-calcification of Bovine Pericardium for Bioprosthetic Heart Valves after Surface Modification with Hyaluronic Acid Derivatives

  • Hahn Sei Kwang;Ohri Rachit;Giachelli Cecilia M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.218-224
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    • 2005
  • Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.

Zero-Length Crosslinking Study on Interactions of TBP, GAL4-AH, and TFIIB in the Preinitiation Complex (전사개시전 복합체에서 TBP, GAL4-AH, TFIIB의 상호작용에 대한 Zero-Length Crosslinking 실험)

  • 권혁만
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.393-399
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    • 1996
  • A zero-Iength croessinking procedure for studying protein-protein interactlons in preinitiation complex has heen developed. Preinltiadon complexes were assembled with immobilized DNA templates coupled to metal beads. Pudfied complexes were dIrectly crosslinked by 1-ethyl-3.(3-dimethylaminopropyl)carbodlimide (EDC). The reaction was stopped by addition of $\beta$-mercaptoethanol, and the complexes were isolated from EDC immedIately. An appllcatlion of this method with a preinltlation complex assembled with TBP, TFIIB, and GAL4-AH demonstrated that ThP dIrectly interacts with GAL4-AH and TFIIB in the prelnitlatlon complex. However, croeslinked produd between GAL4-AH and ThilB was not observed. These resutts lndlcate that GAL4-AH does not stably Interact with TFIIB In the GAL4-AH-TFIIB-TBP-DNA prelnitlatlon complex.

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Electrophoretical Properties of Transglutaminase Treated Milk Product Powders (Transglutaminase를 처리한 분말 유제품의 전기영동적 특성)

  • Jeong, Ji-Eun;Hong, Youn-Ho
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.304-308
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    • 2006
  • This study was performed to understand the behavior of protein mobility and intensity of enzymatic hydrolysis according to crosslinking of sodium caseinate, whey protein isolate, skim milk and whole milk powders with or without transglutaminase (TGase, w/w = 200 : 1) at $38^{\circ}C$. Whey protein was limited to crosslinking and skim milk was relatively more increased in high molecular polymer than whole milk. The degree of crosslinking decreased in the order of sodium caseinate>skim milk>whole milk>whey protein isolate. The SDS-PAGE results indicated that main bands of TGase treated samples had a high mobility and formed bands of molecular weights below 15 kDa by hydrolysis with pepsin after 10 min of reaction time. However, ${\beta}-lactoglobulin$ showed remarkable stability against pepsin hydrolysis treated with and without TGase. The high molecular polymers were easily hydrolyzed with digestive enzymes in vitro experiment. These results suggested that novel dairy products using TGase would have no special digestive problem in human body.

PREVENTION OF HYDROXYL RADICAL-INDUCED ERYTHROCYTE HEMOLYSIS BY PROTEIN THIOLS

  • Youn, Hong-Duk;Packer, Lester;Matsugo, Seiichi
    • Journal of Photoscience
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    • v.4 no.3
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    • pp.133-140
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    • 1997
  • A system for studying oxidative hemolysis has been used by controling UV-irradiation and concentration of a novel molecular probe, N,N'-bis(2-hydroperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetra-carboxylic diimide (NP-III), which generates hydroxyl radical upon longer wavelength photoirradiation (> 350 nm). NP-III induces 25~30% of hemolysis at low concentration (50 $\mu$M) for 3h-irradiation of UVA. The simultaneous treatment of N-ethylmaleimide (NEM) with NP-IH completely hemotyzed erythrocytes under the same conditions as NP-III alone by both decreasing thiol group and increasing lipid peroxidation in erythrocyte membrane. However. thiol-reducing agents prevented the protein-crosslinking and lipid peroxidation on the NEM-synergistic hemolysis by partially scavenging hydroxyl radical and maintaining the thiol group of erythrocyte membrane in the reduced state. In addition, erythrocytes pretreated with 2,2,5,7,8-pentamethyl-6-hydroxychromane (PMC), vitamin E homologue was able to delay and decrease the lipid peroxidation when compared to cells pretreated with both NEM and PMC. We suggest that the presence of reduced thiols in inner membrane protein by GSH can prevent the protein-crosslinking and the lipid peroxidation, and eventually prevent the oxidative hemolysis of erythrocyte.

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Gelation of Rapeseed Protein Induced with Microbial Transglutaminase (미생물성 Transglutaminase에 의한 유채단백질의 겔화)

  • Hyun, Eun-Hee;Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1262-1267
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    • 1999
  • Optimum reaction conditions for gel formation of rapeseed, Brassica napus, protein catalyzed by microbial TGase(transglutaminase) were evaluated by measuring breaking strength and deformation of gel. The polymerization of the protein gel was ascertained by SDS-PAGE and content of GL crosslinking$[{\varepsilon}-({\gamma}-glutamyl)lysine]$. In the reaction between rapeseed protein and TGase at $45^{\circ}C$ for 60 min, the breaking strength and deformation of the gel was the maximum at the ratio of 1 : 40 of enzyme to substrate. 10%(w/v) of rapeseed protein concentrate was optimum for gel production. The maximum breaking strength and deformation was shown at $45^{\circ}C$ The breaking strength increased linearly up to 90 min of the reaction time and remained unchanged. The breaking strength and deformation by TGase treatment was pH dependent and pH 7 was optimum for 10% rapeseed protein solution. SDS-PAGE analysis indicated that new band of highmolecular polymers were formed by the enzyme reaction, with disappearing the original bands of rapeseed protein. According to HPLC analysis. the content of GL crosslinking was increased from 0 to $7.14\;{\mu}mol/g$ gel for 90 min of the reaction time.

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Effect of biofibers addition on the structure and properties of soy protein composite films

  • Ye Eun Kim;Su Jin Kim;Yong-Il Chung;Chae Hwa, Kim;Tae Hee Kim;In Chul Um
    • International Journal of Industrial Entomology and Biomaterials
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    • v.48 no.1
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    • pp.25-32
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    • 2024
  • Soy protein isolate (SPI) has garnered researchers' attention due to its abundance, costeffectiveness, excellent biocompatibility, hemo-compatibility, and biodegradability. However, SPI faces limitations in application due to poor processability and weak mechanical strength. Substantial efforts have been made to address these challenges. In this preliminary study, glycerol and biofibers were added to SPI to improve the mechanical properties and film forming, and glyoxal was employed to crosslink SPI molecules. The microstructure and mechanical properties of the resulting SPI/composite films were evaluated. A 15% addition of glycerol proved sufficient for good film formation. Among the biofibers, short SF microfibers were the most effective in enhancing breaking strength, while TEMPO-oxidized CNF (cellulose nanofiber) excelled among CNFs. Crosslinking with glyoxal significantly enhanced the mechanical properties, with the type of biofiber minimally affecting the mechanical properties of the crosslinked SPI composite films.

Identification of Nuclear Factors that UV-crosslink to Rev-responsive Element RNA (UV조사에 의해 Rev-responsive element RNA와 결합하는 핵단백질인자의 확인)

  • 박희성;남용석
    • Journal of Life Science
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    • v.7 no.3
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    • pp.161-166
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    • 1997
  • HIV-1 Rev protein plays an important role in regulating the expression of viral structural proteins. It allows the nuclear export and accumulation of unspliced and partially spliced viral mRNA in the cytoplasm. The Rev-responsive element RNA, present in the env gene, forms a higly ordered RNA secondary structure and is required for the Rev-mediated mRNA export. For this process to complete factor(s) are strongly suggested. From our experiments of electrophoretic mobility shift, UV-crosslinking and SDS/PAGE, RRE RNA was found to be recognized to several nuclear factors such as 36/37, 56, 41. 76, 150 kD proteins in the order of reactivity. Among them, 36/37 and 56 kD proteins are more reactive upon a brief UV treatment (5 min) and more persistent in the presence of high amount of nonspecific competitor, heparin. Certain nuclear protein9s) seemed to recognize the RRE RNA structure in competition with Rev to gel mobility shift assay.

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Bone Health and L-ascorbic acid in Postmenopausal Women (폐경 여성의 골 건강과 L-ascorbic acid)

  • Kim, Bokyung;Kim, Mihyang
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1142-1148
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    • 2021
  • As the average human lifespan has been extended, there has been a lot of interest in the quality of life of women after menopause. It is known that the average age of menopause among Korean women is 49.7 years, and the post-menopausal life of a woman takes up more than one third of her life. L-ascorbic acid (AsA) is known to be involved in the synthesis and maturation of collagen, a bone constituent protein. The aim of this review is to discuss the potential of AsA in bone health in postmenopausal women. AsA plays an important role in collagen biosynthesis, and collagen is a protein constituting bone and is a necessary material for calcification of the bone matrix. Collagen crosslinking is necessary for the stabilization and elasticity of collagen fibers during growth and matruation of animals, but an excessive increase is likely to lead to further aging because the movement of intercellular nutrients or waste is suppressed. AsA acts as a reducing agent to stabilize the immature collagen crosslinking and suppress pyridinoline production, a mature crosslinking. Therefore, AsA participates in collagen biosynthesis and helps bone tissue health, while regulating the excessive maturation of collagen crosslinking, it is expected to play an important role in bone-related problems that may occur in postmenopausal women.

Denaturation of Heat Treated Lysozyme under High Pressure Conditions (고압조건에서 가열 처리된 Iysozyme의 변성)

  • Cho, Rae-Kwang;Hong, Jin-Hwan
    • Korean Journal of Food Science and Technology
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    • v.23 no.3
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    • pp.366-369
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    • 1991
  • In order to elucidate texturization mechanism of extrudated protein, egg white lysozyme was heated under high pressure conditions, and its solubility and changes of molecular weight were investigated. Under high pressure conditions of $100,\;300\;and\;600\;kg/cm^2$, solubility decreased gradually with increasing temperature in the samples heated at $70,\;120\;and\;150^{\circ}C$ and decreased notably with increasing pressure at $200^{\circ}C$. Polymerization was found in the samples heated at $150\;and\;200^{\circ}C$ while a band which located below monomer(low-molecular) could be recognized. Molecular weight of the low-molecular was estimated to be about $6,000{\sim}9,000$ and no smaller peptide was recognized. The polymerization may have occured by disulfide crosslinking in the samples heated at $120^{\circ}C$ but other crosslinking may have played a role in those at $150\;and\;200^{\circ}C$.

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Bio-film Composites Composed of Soy Protein Isolate and Silk Fiber: Effect of Concentration of Silk Fiber on Mechanical and Thermal Properties

  • Prabhakar, M.N.;Song, Jung Il
    • Composites Research
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    • v.27 no.5
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    • pp.196-200
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    • 2014
  • A novel, simple and totally recyclable method has been developed for the synthesis of nontoxic, biocompatible and biodegradable bio-composite films from soy protein and silk protein. Bio films are defined as flexible films prepared from biological materials such as protein. These materials have potential application in medical and food as a packaging material. Their use depends on various parameters such as mechanical (strength and modulus), thermal, among others. In this study, prepare and characterization of bio films made from Soy Protein Isolate (SPI) (matrix) and Silk Fiber (SF) (reinforcement) through solution casting method by the addition of plasticizer and crosslinking agent. The obtained SPI and SPI/SF composites were subsequently subjected to evaluate their mechanical and thermal properties by using Universal Testing Machine and Thermal Gravimetric Analyzer respectively. The tensile testing showed significant improvements in strength with increasing amount of SF content and the % elongation at break of the composites of the SPI/SF was lower than that of the matrix. Though the interfacial bonding was moderate, the improvement in tensile strength and modulus was attributed to the higher tensile properties of the silk fiber.