Advanced
Rod and Vesicular Structures of Cyclosophoraose-Based Ionic Self-assembly
Rod and Vesicular Structures of Cyclosophoraose-Based Ionic Self-assembly
Bulletin of the Korean Chemical Society. 2014. Aug, 35(8): 2537-2540
Copyright © 2014, Korea Chemical Society
  • Received : March 15, 2014
  • Accepted : April 05, 2014
  • Published : August 20, 2014
Download
PDF
e-PUB
PubReader
PPT
Export by style
Article
Author
Metrics
Cited by
TagCloud
About the Authors
Eunae Cho
Daham Jeong
Seung R. Paik
Department of Chemical & Biological Engineering, Seoul National University, Seoul 151-742, Korea
Seunho Jung

Abstract
Keywords
PPT Slide
Lager Image
PPT Slide
Lager Image
PPT Slide
Lager Image
Experimental
Purification of Cys. The isolation and purification of Cys from Rhizobium trifolii TA-1 were carried out as previously described. 26 Culture supernatants were concentrated fivefold by rotary evaporation, and the concentrated sample was precipitated by adding 3 volumes of ethanol. After centrifugation, the supernatant was concentrated by rotary evaporation and the product was collected by adding 7 volumes of ethanol. After decanting the supernatant, the precipitates were applied to Bio-Gel P-6 column. The fractions were assayed for carbohydrates using the phenol-sulfuric acid method. The fractions that contained Cys were pooled, concentrated, and desalted using Bio-Gel P-4 column. The desalted Cys were confirmed by NMR spectroscopy and MALDI-TOF MS.
Synthesis of QA–Cys. Cys (1 g) was dissolved in 7% NaOH aqueous solution for 30 min, and 2,3-epoxypropyltrimethylammonium chloride (3.1 g) was added. The mixture was reacted for 5 h at 65 °C, neutralized with HCl and evaporated to a viscous residue. After desalting on a Bio-gel P-2 column, the product was lyophilized, and the final structure was elucidated with EA (ThermoFinnigan, Flash2000) and NMR spectroscopy.
Synthesis of CM–Cys. CM–Cys was prepared as described in a previous report. 6 Cys (500 mg) and NaOH (2.8 g) were mixed in distilled water (7.4 mL). Monochloroacetic acid solution (20%) was added. After reacting for 4 h at 50 °C, the mixture was neutralized with 6 M HCl. The mixture was precipitated with 5 vol MeOH and left overnight at 4 °C. The precipitate was dissolved in water and desalted on a Sephadex G-10 column. The product was confirmed using MALDI-TOF MS (Voyager-DETM STR Biospectrometry Workstation) and NMR spectroscopy.
Nuclear Magnetic Resonance (NMR) Spectroscopy. For the NMR spectroscopic analysis, a Bruker Avance 500 spectrometer was used to record the 1 H–NMR, 1 C-NMR, DEPT and HSQC spectra. The HSQC spectrum was measured with a spectral width of 3401 Hz in both dimensions and 256/2048 complex data points in t1 and t2 , respectively. NMR analyses were performed in D 2 O at room temperature.
Scanning Electron Microscopy (SEM). QA–Cys (3 mM) and CM–Cys (3 mM) were mixed in 100 μL of 20 mM phosphate buffer (pH 7). Hexane was added to a QA–Cys/ CM–Cys mixture in phosphate buffer (pH 7), and a turbid solution was obtained by vortexing for 5 min. After centrifugation, the precipitate was lyophilized. The lyophilized samples were mounted onto stubs using double-sided adhesive tape and then made electrically conductive by coating with a thin layer of gold. The surface morphologies of the materials were examined under a scanning electron microscope (Jeol, JSM 6380, Tokyo, Japan).
Transmission Electron Microscopy (TEM). QA–Cys (3 mM) and DCA (3 mM; Sigma Aldrich) were mixed in 800 μL of 20 mM phosphate buffer (pH 7). After sonication for 30 s, the aqueous suspension (10 μL) containing the supramolecular aggregates formed by the QA–Cys/DCA mixture was adsorbed onto a carbon-coated copper grid (300-mesh) and air-dried for 1 min. For clear negative staining, the supernatant of 2% uranyl acetate following centrifugation at 13,200 rpm for 2 min was used. The aggregates were examined using a transmission electron microscope (JEOL, JEM 1010, Tokyo, Japan).
Dynamic Light Scattering (DLS). DLS measurements were carried out with a Wyatt Technology DynaPro Plate Reader at constant room temperature.
Acknowledgements
This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2013R1A1A2012568 and NRF- 2011-619-E0002) and supported by the Priority Research Centers Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2012-0006686). SDG.
References
Breedveld M. W. , Miller K. J. 1994 Microbiol. Rev. 58 145 -
Andre’ L. , Mazeau K. , Taravel F. R. , Tvaroska I. 1995 Int. J. Biol. Macromol. 17 189 -    DOI : 10.1016/0141-8130(95)92685-J
Koizumi K. , Okada Y. , Horiyama S. , Utamura T. , Higashiura T. , Ikeda M. 1984 J. Incl. Phenom. 2 891 -    DOI : 10.1007/BF00662259
Lee S. , Seo D. , Kim H. W. , Jung S. 2001 Carbohydr. Res. 334 119 -    DOI : 10.1016/S0008-6215(01)00178-1
Piao J. , Jang A. , Choi Y. , Tahir M. N. , Kim Y. , Park S. , Cho E. , Jung S. 2014 Carbohydr. Polym. 101 733 -    DOI : 10.1016/j.carbpol.2013.09.104
Lee S. , Park H. , Seo D. , Choi Y. , Jung S. 2004 Carbohydr. Res. 339 519 -    DOI : 10.1016/j.carres.2003.11.011
Sun T. , Guo Q. , Zhang C. , Hao J. , Xing P. , Su J. , Li S. , Hao A. , Liu G. 2012 Langmuir 28 8625 -    DOI : 10.1021/la301497t
Zhang H. , Liu Z. , Xin F. , An W. , Hao A. , Li J. , Li Y. , Sun L. , Sun T. , Zhao W. , Li Y. , Kong L. 2011 Carbohydr. Res. 346 294 -    DOI : 10.1016/j.carres.2010.11.010
Lehn J. M. 2002 Science 295 2400 -    DOI : 10.1126/science.1071063
Faul C. F. J. , Antonietti M. 2003 Adv. Mater. 15 673 -    DOI : 10.1002/adma.200300379
Xu J. , Bai H. , Yi C. , Luo J. , Yang C. , Xia W. , Liu X. 2011 Carbohydr. Polym. 86 678 -    DOI : 10.1016/j.carbpol.2011.05.006
Kujawa P. , Moraille P. , Sanchez J. , Badia A. , Winnik F. M. 2005 J. Am. Chem. Soc. 127 9224 -    DOI : 10.1021/ja044385n
Costalat M. , David L. , Delair T. 2014 Carbohydr. Polym. 102 717 -    DOI : 10.1016/j.carbpol.2013.10.098
Fan L. , Cao M. , Gao S. , Wang W. , Peng K. , Tan C. , Wen F. , Tao S. , Xie W. 2012 Carbohydr. Polym. 88 707 -    DOI : 10.1016/j.carbpol.2012.01.021
Balasubramanian D. , Raman B. , Sundari C. S. 1993 J. Am. Chem. Soc. 115 74 -    DOI : 10.1021/ja00054a010
Das K. , Sarkar N. , Das S. , Bhattacharyya K. 1995 Langmuir 11 2410 -    DOI : 10.1021/la00007a016
Steed J. W. , Atwood J. L 2009 Supramolecular Chemistry 2nd ed. John Wiley & Sons, Inc. New York, USA
Lee J. , Bhak G. , Lee S. , Paik S. R. 2008 Biophys. J. L16 -
Venneman N. G. , van Kammen M. , Renooij W. , van Berge- Henegouwen G. P. , van Erpecum K. J. 2005 Biochim. Biophys. Acta 1686 209 -    DOI : 10.1016/j.bbalip.2004.10.004
Sun T. , Zhang J. , Kong L. , Qiao H. , Li Y. , Xin F. , Hao A. 2011 Carbohydr. Res. 346 285 -    DOI : 10.1016/j.carres.2010.11.003
Kim Y. H. , Gihm S. H. , Park C. R. 2001 Bioconjugate Chem. 12 932 -    DOI : 10.1021/bc015510c
Kanemaru M. , Yamamoto K. , Kadokawa J. 2012 Carbohydr. Res. 357 32 -    DOI : 10.1016/j.carres.2012.05.014
Allen T. M. , Cullis P. R. 2004 Science 303 1818 -    DOI : 10.1126/science.1095833
Christense S. M. , Stamou D. 2007 Soft Matter 3 828 -    DOI : 10.1039/b702849k
Li J. H. , Wang Y. F. , Ha W. , Liu Y. , Ding L. S. , Li B. J. , Zhang S. 2013 Biomacromolecules 14 2984 - 2988    DOI : 10.1021/bm400584h
Jeon Y. , Kwon C. , Cho E. , Jung S. 2010 Carbohydr. Res. 345 2408 -    DOI : 10.1016/j.carres.2010.08.009