• Title/Summary/Keyword: Peptide conjugate

Search Result 19, Processing Time 0.024 seconds

Low Molecular Weight Polyethylenimine-Mitochondrial Leader Peptide Conjugate for DNA Delivery to Mitochondria

  • Choi, Joon-Sig;Choi, Min-Ji;Go, Gyeong-Su;Rhee, Byoung-Doo;KimPak, Young-Mi;Bang, In-Seok;Lee, Min-Hyung
    • Bulletin of the Korean Chemical Society
    • /
    • v.27 no.9
    • /
    • pp.1335-1340
    • /
    • 2006
  • It has been found that a number of diseases are associated with mutations in the mitochondrial DNA. Therapeutic gene delivery to mitochondria has been suggested as a clinical option for these diseases. In this study, we developed a gene carrier to mitochondria by the conjugation of mitochondrial leader peptide (LP) to polyethylenimine (PEI). Mitochondrial LP conjugated PEI (PEI-LP) was synthesized with low molecular weight PEI (2,000 Da, PEI2K). Gel retardation assay showed that PEI2K-LP formed complexes at a 1.0/1 weight ratio. In addition, PEI2K-LP protected DNA from the enzymatic degradation for at least 60 min, while naked DNA was completely degraded within 20 min. PEI2K-LP was compared with LP conjugated high molecular weight PEI (25,000 Da, PEI25K) in terms of toxicity and delivery efficiency. MTT assay showed that PEI2K-LP had much lower cytotoxicity than PEI25K-LP to 293 cells. In addition, cell-free DNA delivery assay showed that PEI2K-LP delivered more DNA to mitochondria at a 1.8/1 weight ratio than naked DNA or PEI. This result suggests that PEI2K-LP may be useful for the development of mitochondrial gene therapy system with lower cytotoxicity.

Specific Targeting of Fluorescein Isothiocyanate with Ep-CAM Antibody(Specific targeting of FITC with Ep-CAM Antibody)

  • Lee, Young-Tae;Tae, Gun-Sik
    • Journal of Photoscience
    • /
    • v.10 no.3
    • /
    • pp.237-240
    • /
    • 2003
  • The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

  • PDF

Conjugation of Protein and Peptide Drugs with Hydrophilic Polymers and Their Applications (수용성 고분자물질-단백질 접합체의 합성 및 응용)

  • Yong, Chul-Soon;Sohn, Young-Taek
    • Journal of Pharmaceutical Investigation
    • /
    • v.23 no.4
    • /
    • pp.187-206
    • /
    • 1993
  • Since the advent of recombinant DNA technology coupled with other biotechnology a variety of therapeutically effective proteins and peptides have been extensively invesitigated and many of them are now on clinical trial. They, however, suffer from some problems such as immunogenicity, antigenicity, instability and short half-life in circulation due to their proteinous natures. These drawbacks can be overcome successfully by conjugating proteins and peptides with hydrophilic polymers such as polyethylene glycol (PEG), albumin or dextran. The resulting soluble conjugates showed reduced antigenicity and immunogenicity, increased circulatory half-life, enhanced stability against proteolytic degradation. Comparing with the unmodified proteins and peptides, the therapeutic potential of conjugates is greatly enhanced. Clinical applications of these conjugates have shown promising results for the future use.

  • PDF

Studies on the Selectivity of Herbicide Alachlor;I. Phytotoxicity and Glutathione Conjugation (제초제 Alachlor의 선택성에 관한 연구;I. 약해와 글루타치온 Conjugation 반응)

  • Park, Chang-Kyu;Hwang, Eul-Chul
    • Korean Journal of Environmental Agriculture
    • /
    • v.6 no.1
    • /
    • pp.44-49
    • /
    • 1987
  • Present work has been initiated to see if inherent biochemical difference among plants is, in any way, related to the observed selectivity characteristics of preemergence herbicide, alachlor. Application of aqueous solution of alachlor onto three intact plants, soybean, chinese cabbage and barnyard grass resulted in phytotoxicity responses in the testt plants in varying degree. Examination of glutathione (and homoglutathione) contents of the test plants indicated that the phytotoxicity is inversely proportional to the peptide contents of the test plants. It was also noted that four to five water soluble metabolites are readily formed in intact seedling treated with labelled alachlor and glutathionealachlor and homoglutathionealachlor conjugates were tentatatively identified as major metabolites. It is concluded that conjugation reaction involving glutathiones and xenobiotic alachlor, a typical phase II reaction, acts as detoxification reaction in the three test plants and this would, in turn, contribute to observed selectivity of alachlor.

  • PDF

Cell-Specific Targeting of Texas Red with Anti-Ep-CAM Antibody

  • Lee, Soo-Chul;Tae, Gun-Sik
    • Journal of Photoscience
    • /
    • v.12 no.3
    • /
    • pp.123-127
    • /
    • 2005
  • The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

  • PDF

Cloning of Chicken Microsomal Glutathione S-transferase 1 Gene (MGST1) and Identification of Its Different Splice Variants

  • Wang, X.-T.;Zhang, H.;Zhao, C.-J.;Li, J.-Y.;Xu, G.-Y.;Lian, L.-S.;Wu, C.-X.;Deng, Xuemei
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.22 no.2
    • /
    • pp.155-161
    • /
    • 2009
  • Mammal microsomal glutathione transferase 1 (MGST1) can conjugate many toxic or carcinogenic substances and depress oxidative stress. In this study, Chicken MGST1 and its variants were cloned for the first time and were composed of 956 or 944 nucleotides. The 12 nt deletion in the exon 2 did not alter the GT-AG rule and the ORFs for the two MGST1 variants were the same, which both comprised 465 nucletides and encoded a peptide with 155 amino acids. It was found that the two different splice variants identified using RT-PCR expressed in all three organs investigated of Dwarf Brown Chicken, namely liver, spleen and shell gland. Moreover, the expression level of MGST1 mRNA in the liver of Dwarf Brown chickens was the highest (p<0.01), and there were no significant differences between the spleen and the shell gland. These results provide a base for studying the biological function of Chicken MGST1.

A STUDY ON THE DISTRIBUTION OF CALCITONIN GENE-RELATED PEPTIDE CONTAINING NERVE FIBERS IN RAT PULP FOLLOWING DENTINAL INJURY (상아질 손상 후 흰쥐 대구치 치수의 calcitonin gene-related peptide(CGRP) 함유 신경섬유 분포에 관한 연구)

  • Moon, Joo-Hoon;Park, Sang-Jin;Min, Byung-Soon;Choi, Ho-Young;Cho, Gi-Woon
    • Restorative Dentistry and Endodontics
    • /
    • v.24 no.1
    • /
    • pp.100-115
    • /
    • 1999
  • The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

  • PDF

Stereoselective Synthesis of Novel Bestatin Analogs (새로운 베스타틴 유사체의 입체선택적 합성)

  • Seo, Youngran;Lee, SooBeom;Kim, Young Gyu
    • Applied Chemistry for Engineering
    • /
    • v.26 no.1
    • /
    • pp.111-115
    • /
    • 2015
  • Two new analogs of bestatin were prepared from $\small{D}$-leucine and $\small{D}$-valine in a stereoselective and efficient way. An aminopeptidase inhibitor bestatin shows significant biological effects on immunomodulation and is marketed for the treatment of acute myelocytic leukemia. The key intermediates, trans-oxazolidine methyl esters 2a and 2b, were obtained with more than 20 to 1 stereoselectivity in a one-pot procedure by the three cascade reactions between N-hydroxymethyl protected ${\alpha}$-amino aldehydes (4a and 4b) and phenylsulfonylnitromethane ($PhSO_2CH_2NO_2$) and the following in-situ ozonolysis. Basic hydrolysis of 2a and 2b, and then the peptide coupling with $\small{L}$-Leu-OMe produced the protected derivatives of two new bestatin analogs, 3a and 3b, respectively. The new isobutyl and isopropyl analogs of bestatin (1a and 1b) were produced in overall 51% and 38% yields, respectively, with high stereoselectivity from the corresponding protected ${\alpha}$-amino aldehydes 4 in a six-step process.