• Title/Summary/Keyword: Synthetic peptide

Search Result 205, Processing Time 0.039 seconds

Hydrogel microrheology near the liquid-solid transition

  • Larsen, Travis;Schultz, Kelly;Furst, Eric M.
    • Korea-Australia Rheology Journal
    • /
    • v.20 no.3
    • /
    • pp.165-173
    • /
    • 2008
  • Multiple particle tracking microrheology is used to characterize the viscoelastic properties of biomaterial and synthetic polymer gels near the liquid-solid transition. Probe particles are dispersed in the gel precursors, and their dynamics are measured as a function of the extent of reaction during gel formation. We interpret the dynamics using the generalized Stokes-Einstein relationship (GSER), using a form of the GSER that emphasizes the relationship between the probe particle mean-squared displacement and the material creep compliance. We show that long-standing concepts in gel bulk rheology are applicable to microrheological data, including time-cure superposition to identify the gel point and critical scaling exponents, and the power-law behavior of incipient network's viscoelastic response. These experiments provide valuable insight into the rheology, structure, and kinetics of gelling materials, and are especially powerful for studying the weak incipient networks of dilute gelators, as well as scarce materials, due to the small sample size requirements and rapid data acquisition.

Production and Characterization of Specific Antibodies to Bombesin

  • Kwon, Hyeok-Yil;Lee, Yun-Lyul;Park, Hyoung-Jin
    • The Korean Journal of Physiology
    • /
    • v.28 no.1
    • /
    • pp.91-97
    • /
    • 1994
  • In order to produce a specific bombesin antiserum far very sensitive radioimmunoassay, synthetic $[lys^3]-bombesin$ conjugated to bovine serum albumin was subcutaneously injected into guinea pigs. The conjugation was performed using either carbodiimide or gIutaraldehyde as a coupling agent. The antisera were characterized by analysis of Scatchard and Sips plots. The antiserum LBE 2G/2 raised by repeat injection of the immunogen conjugated with carbodiimide showed the titer of 1 : 188,000, very low cross-reactivity to bombesin-like peptides except bombesin, with high affinity constant $(1.64{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.91). The antiserum LBG 1G/2 produced by repeat injection of the immunogen conjugated with glutaraldehyde possessed the titer of 1 : 43,000, high cross-reactivity to some bombesin-like peptides, high affinity constant $(1.19{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.79). These results indicate that the antiserum LBE 2G/2 is specific only to bombesin and that the antiserum LBG IG/2 binds to some bombesin-like peptides such as alytesin, gastrin releasing peptide and neuromedin C. The antiserum LBE 2G/2 is sufficient for the very sensitive radioimmunoassay of bombesin.

  • PDF

New Materials Based Lab-on-a-Chip Microreactors: New Device for Chemical Process

  • Kim, Dong-Pyo
    • Proceedings of the Materials Research Society of Korea Conference
    • /
    • 2012.05a
    • /
    • pp.51-51
    • /
    • 2012
  • There is a growing interest in innovative chemical synthesis in microreactors owing to high efficiency, selectivity, and yield. In microfluidic systems, the low-volume spatial and temporal control of reactants and products offers a novel method for chemical manipulation and product generation. Glass, silicon, poly(dimethylsiloxane) (PDMS), and plastics have been used for the fabrication of miniaturized devices. However, these materials are not the best due to either of low chemical durability or expensive fabrication costs. In our group, we have recently addressed the demand for economical resistant materials that can be used for easy fabrication of microfluidic systems with reliable durability. We have suggested the use of various specialty polymers such as silicon-based inorganic polymers and fluoropolymer, flexible polyimide (PI) films that have not been used for microfluidic devices, although they have been used for other areas. And inexpensive lithography techniques were used to fabricate Lab-on-a-Chip type of microreactors with differently devised microchannel design. These microreactors were demonstrated for various synthetic reactions: liquid, liquid-gas organic chemical reactions in heterogeneous catalytic processes, syntheses of polymer and non-trivial inorganic materials. The microreactors were inert, and withstand even harsh conditions, including hydrothermal reaction. In addition, various built-in microstructures inside the microchannels, for example Pd decorated peptide nanowires, definitely enhance the uniqueness and performance of microreactors. These user-friendly Lab-on-a-Chip devices are useful alternatives for chemist and chemical engineer to conventional chemical tools such as glass.

  • PDF

Cloning, Expression Analysis and Enzymatic Characterization of Cathepsin L from the Inshore Hagfish (Eptatretus burgeri) (먹장어 Cathepsin L의 분자생물학적 클로닝, 발현 및 효소학적 특성 분석)

  • JANG, Jin-Hyeon;SON, So-Hee;JO, Hyeon-Kyeong;CHUNG, Joon-Ki;LEE, Hyung-Ho
    • Journal of Fisheries and Marine Sciences Education
    • /
    • v.28 no.4
    • /
    • pp.903-912
    • /
    • 2016
  • Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

Antimicrobial activity and characterization for defensin of synthetic oligopeptides derived from Bombus ignitus (호박벌 유래 디펜신 유전자의 분자적 특성분석 및 항균 활성)

  • Kang, Heui-Yun;Kim, In-Woo;Lee, Joon-Ha;Kwon, Young Nam;Yun, Eun-Young;Yoon, Hyung Joo;Kim, Seong-Ryul;Kim, Iksoo;Hwang, Jae-Sam
    • Journal of Sericultural and Entomological Science
    • /
    • v.50 no.2
    • /
    • pp.161-165
    • /
    • 2012
  • Antimicrobial peptides of insects are found and reported as immune defence system against infectious agents. The peptides are produced by fat body cells and thrombocytoids, a blood cell type. Defensin is 38-45 amino acids long and consists of an ${\alpha}$-helix linked by a loop to an antiparallel ${\beta}$-sheet. Defensin from a bumblebee, Bombus ignitus, is known to comprise 52 amino acid residues. This peptide consists of two ${\alpha}$-helixes; ACAANCLSM and KTNFKDLWDKRF and one ${\beta}$-sheet; GGRCENGVCLCR. We carried out antibacterial activity test by radial diffusion assay against Staphylococcus aureus (Gram positive), Escherichia coli (Gram negative), Pseudomonas syringae (Gram negative), Candida albicans (fungi), MDRPA, MRSA, and VRE (antimicrobial resistant microbes) with synthetic oligopeptides from Peptron (Daejeon, Korea). The predicted curtailment fragment (GGRCEVCLCR-$NH_2$) for ${\beta}$-sheet had strong antibacterial activity when internal amino acids were removed. But, curtailment fragments (ACAANCLSM-$NH_2$ and TNFKDLWDKR-$NH_2$) of ${\alpha}$-helix were not showed antibacterial activity. These synthetic oligopeptides were showed the great activity against Gram positive and negative bacteria.

Isolation of Bombesin-Like Substances from the Skin of the Frog, Bombina orientalis: Its Molecular Heterogeneity and Biological Activity (한국에 서식하는 무당개구리의 피부에서 추출된 Bombesin 유사물질의 분자적 이질성 및 생물학적 활성)

  • Park, Hyoung-Jin;Lee, Yun-Lyul;Kwon, Hyeok-Yil;Shin, Won-Im;Suh, Sang-Won
    • The Korean Journal of Physiology
    • /
    • v.23 no.1
    • /
    • pp.79-87
    • /
    • 1989
  • Molecular property as well as biological activities of bombesin-like substance (BBS-LS) isolated from the skin of B. orientalis in Korea was compared with those of synthetic BBS-14. BBS-LS in the crude extract was applied on a Sephadex G-50, superfine column $(1.6{\times}112\;cm)$. On chromatogram, the first peak (3.5% of BBS-LS) was found ahead of synthetic GRP-27, and the second peak (rest) between synthetic GRP-27 and synthetic BBS-14. The main form of BBS-LS was successfully purified by using a column of alkaline alumina followed by sequential gel-filtrations on a column of Sephadex G-10, fine and a column of Sephadex G-50, superfine. Chromatographic analysis of the purified BBS-LS using a column of Sephadex G-50, superfine and reversed phase HPLC revealed that the main form of BBS-LS in the skin of B. orientalis could be distinctly different from either BBS-14 or GRP-27 in molecular size. The purified BBS-LS exerted biological activities quite identical to those of synthetic BBS-14. The results of the present investigation indicate that the skin of B. orientalis contains BBS-LS composed of two distinct forms. The main form of BBS-LS purified in the present study is heterogenous to both synthetic BBS-14 and GRP-27 in molecular size but identical to BBS in biological activities.

  • PDF

Aquatide Activation of SIRT1 Reduces Cellular Senescence through a SIRT1-FOXO1-Autophagy Axis

  • Lim, Chae Jin;Lee, Yong-Moon;Kang, Seung Goo;Lim, Hyung W.;Shin, Kyong-Oh;Jeong, Se Kyoo;Huh, Yang Hoon;Choi, Suin;Kor, Myungho;Seo, Ho Seong;Park, Byeong Deog;Park, Keedon;Ahn, Jeong Keun;Uchida, Yoshikazu;Park, Kyungho
    • Biomolecules & Therapeutics
    • /
    • v.25 no.5
    • /
    • pp.511-518
    • /
    • 2017
  • Ultraviolet (UV) irradiation is a relevant environment factor to induce cellular senescence and photoaging. Both autophagy- and silent information regulator T1 (SIRT1)-dependent pathways are critical cellular processes of not only maintaining normal cellular functions, but also protecting cellular senescence in skin exposed to UV irradiation. In the present studies, we investigated whether modulation of autophagy induction using a novel synthetic SIRT1 activator, heptasodium hexacarboxymethyl dipeptide-12 (named as Aquatide), suppresses the UVB irradiation-induced skin aging. Treatment with Aquatide directly activates SIRT1 and stimulates autophagy induction in cultured human dermal fibroblasts. Next, we found that Aquatide-mediated activation of SIRT1 increases autophagy induction via deacetylation of forkhead box class O (FOXO) 1. Finally, UVB irradiation-induced cellular senescence measured by $SA-{\beta}-gal$ staining was significantly decreased in cells treated with Aquatide in parallel to occurring SIRT1 activation-dependent autophagy. Together, Aquatide modulates autophagy through SIRT1 activation, contributing to suppression of skin aging caused by UV irradiation.

Study of Anti-inflammatory Effect of CopA3 Peptide Derived from Copris tripartitus (애기뿔소똥구리 유래 CopA3합성 펩타이드의 항염증 효능에 관한 연구)

  • Kim, Hyeon-Jeong;Kim, Dong-Hee;Lee, Jin-Young;Hwang, Jae-Sam;Lee, Joon-Ha;Lee, Seul-Gi;Jeong, Hyeon-Guk;An, Bong-Jeun
    • Journal of Life Science
    • /
    • v.23 no.1
    • /
    • pp.38-43
    • /
    • 2013
  • The objective of this study was to evaluate the effect of the synthetic CopA3 peptide of Copris tripartitus on skin inflammation. Regulatory mechanisms of cytokines and nitric oxide (NO) are involved in the immunological activity of RAW 264.7 cells. Tested cells were treated with different concentrations of CopA3 and further cultured for an appropriate time after lipopolyssacharide (LPS) addition. During the entire experimental period, 5, 25, 50, and 100 ${\mu}g/ml$ of CopA3 had no cytotoxicity. At these concentrations, CopA3 inhibited tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). CopA3 also inhibited the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). CopA3 inhibited the activity of iNOS and COX-2 by 41% and 59%, respectively, at 100 ${\mu}g/ml$. In addition, CopA3 reduced the release of inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These results suggest that CopA3 may have significant effects on inflammatory factors and that it may be a potential anti-inflammatory therapeutic agent.

Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52 (Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현)

  • Joo, Han-Seung;Choi, Jang Won
    • Journal of agriculture & life science
    • /
    • v.45 no.6
    • /
    • pp.201-212
    • /
    • 2011
  • The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.

Hyaluronic Acid Enhances the Dermal Delivery of Anti-wrinkle Peptide via Increase of Stratum Corneum Fluidity (히알루론산의 각질 유동성 향상을 통한 주름 개선 펩타이드 피부 흡수 촉진)

  • Kim, Yun-Sun;Kim, Daehyun;Kim, Yumi;Park, Sun-Gyoo;Lee, Cheon-Koo;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.44 no.4
    • /
    • pp.447-453
    • /
    • 2018
  • Acetyl hexapeptide 8 (AH8) is a synthetic peptide for anti-wrinkle cosmetics ingredient. It was developed as a mimetic of botox, patternd after N -terminal end of the protein synatosomal-associated protein 25 (SNAP25), a substrate of botulinum toxin. While AH8 has good efficacy and safety profiles, the permeation through the skin is poor. Therefore, we tried to enhance the transdermal delivery of AH8 by using of hyaluonic acid (HA), a linear polysaccharide of N-acetyl glucosamine and glucuronic acid. To investigate the effect of HA on AH8 penetration, we analyzed paraffin sections of $Micropig^{(R)}$ skin. Fluorescence labeled AH8 was applied to micropig skin with or without HA. The absorption of AH8 was limited to the stratum corneum (SC) without HA. On the other hand, AH8 penetrated to the dermis with HA. Especially, low molecular weight HA (5 kDa) was most efficient compared to 500 kDa HA and 2000 kDa HA. Experiments using fourier-transform infrared (FTIR) spectroscopy revealed that lower molecular weight HA had a tendency to increase the fluidity of the SC lipids more, which means enhancing the skin penetration. Therefore, HA could be expected to enhance the anti-wrinkle effect of AH8.