• Title/Summary/Keyword: Peptide-Binding

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Production of polyclonal anti-$\beta$-adrenergic receptor antibody and it′s effects on receptor ligand binding

  • Kim, Hee-Jin;Shin, Chan-Young;Noh, Min-Su;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.86-86
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    • 1995
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently by the use of specific anti-receptor antibodies. A 14-mer peptide (from Phe102 to Leu115 of ${\beta}$2-adrenergic receptor) was synthesized and this peptide was coupled to carrier protein Keyhole Limpet Hemocyanin(KLH) by glutaraldehyde method. A 0.5mg of KLH-coupled peptide was emulsified with equal volume of complete Freund's adjuvant and injected via popliteal lymph node to each of the three Newzealnd White rabbits. Booster injections were repeated at 4 weeks interval for three times with incomplete Freund's adjuvants. One week after the final injection, serum was prepared from ear artery. Nonspecific immunoglobulins were removed by passing the serum through KLH-Sepharose 6B affinity matrix and further by incubation with bovine lung aceton powder. The titer of the antibody for synthetic peptide which was determined by enzyme linked immunosorbent assay(ELISA) was about l/l,000. The antibody produced in this study revealed 67kDa protein band in the western blot of partially purified guinea pig lung ${\beta}$-adrenergic receptor preparation. The antibody inhibited ${\beta}$-adrenergic antaginist [3H] Dihydroalprenolol binding to soluble ${\beta}$-adrenergic receptor by 25% while control sera did not show any inhibitory effects, The result of this study suggests that the peptide sequence selected in this study may play some important roles in adrenergic receptor-ligand interaction.

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The Spectroscopic Study on the Role of C-terminal Region of T4 endonuclease V in the Interaction with DNA: NMR and Fluorescence Experiment (DNA와 상호작용에서 T4 endonuclease V의 C-말단 부위의 역할에 관한 분광학적 연구: 핵자기공명과 형광 실험)

  • Yu, Jun-Seok;Lihm, Hyung-Mi;Ihm, Hu-Kang;Shin, Jung-Hyu;Lee, Bong-Jin
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.193-201
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    • 1996
  • In order to study the role of C-terminal aromatic region of T4 endonuclease V in the interaction with substrate DNA, NMR and Fluorescence spectrum were recorded. Analysis of flu orescence emission spectra showed that C-terminal region of T4 endonuclease V is in or very near the binding site. In the HSQC spectrum of $^{15}N$-Tyr-labeled T4 endonuclease V*DNA complex, the broadening of a peak was observed. It is presumed that this peak corresponds to one among three tyrosine residues which belong to the WYKYY segment of C-terminal region of T4 endonuclease V. Interactions of peptide fragments consisting of C-terminal residues of T4 endonuclease V with DNAs(TT-, T^T-DNA) were investigated by NMR and Fluorescence experiment. The results suggest that two peptide fragments themselves bind to DNAs and their binding pattern is not an intercalation mode.

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Isolation of calcium-binding peptides from porcine meat and bone meal and mussel protein hydrolysates (돼지 육골분 및 진주담치 단백질의 가수분해물 제조 및 칼슘 결합 물질의 분리)

  • Jung, Seung Hun;Song, Kyung Bin
    • Food Science and Preservation
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    • v.22 no.2
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    • pp.297-302
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    • 2015
  • Calcium is one of the essential mineral for the humans due to its crucial physiological functions in the body. Calcium deficiency results in many diseases, such as osteoporosis. Therefore, calcium supplements are available as a functional food. However, most calcium supplements in the market have a limitation due to poor absorption and low bioavailability. Thus, calcium-chelated peptides for improving the absorption rate of calcium have been isolated from foods including porcine meat and bone meal (MBM), and mussel using the enzymatic hydrolysis of their protein. The hydrolysates of food were ultra-filtered in order to obtain small peptides less than 3 kDa and the Ca-binding peptides were isolated via the anion exchange chromatography. The binding activity and concentration of Ca-binding pepetides were determined. In particular, the MBM and mussel protein hydrolysates were fractionated by mono Q and Q-Sepharose, respectively. As a result, among the fractions, the fractions of MBM F2 and mussel F3 showed the highest Ca-binding activity. These results suggest that MBM and mussel protein hydrolysates can be used as calcium supplements.

Immobile Artificial Metalloproteases

  • Kim, Myoung-Soon;Suh, Jung-Hun
    • Bulletin of the Korean Chemical Society
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    • v.26 no.12
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    • pp.1911-1920
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    • 2005
  • Effective artificial metalloproteases have been designed by using cross-linked polystyrene as the backbone. Artificial active sites comprising Cu(II) complexes as the catalytic site and other metal centers or organic functionalities as binding sites were synthesized. The activity of Cu(II) centers for peptide hydrolysis was greatly enhanced on attachment to polystyrene. By placing binding sites in proximity to the catalytic centers, the ability to hydrolyze a variety of protein substrates at selected cleavage sites was improved. Thus far, the most advanced immobile artificial proteases have been obtained by attaching the aldehyde group in proximity to the Cu(II) complex of cyclen.

Thermodynamics of Partitioning of Substance P in Isotropic Acidic Bicelles

  • Baek, Seung Bin;Lee, Hyeong Ju;Lee, Hee Cheon;Kim, Chul
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.743-748
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    • 2013
  • The temperature dependence of the partition coefficients of a neuropeptide, substance P (SP), in isotropic acidic bicelles was investigated by using a pulsed field gradient nuclear magnetic resonance diffusion technique. The addition of negatively charged dimyristoylphosphatidylserine to the neutral bicelle changed the SP partitioning a little, which implies that the hydrophobic interaction between the hydrophobic residues of SP and the acyl chains of lipid molecules is the major interaction while the electrostatic interaction is minor in SP binding in a lipid membrane. From the temperature dependence of the partition coefficients, thermodynamic functions were calculated. The partitioning of SP into the acidic bicelles is enthalpy-driven, as it is for small unilamellar vesicles and dodecylphosphocholine micelles, while peptide partitioning into a large unilamellar vesicle is entropy-driven. This may mean that the size of lipid membranes is a more important factor for peptide binding than the surface curvature and surface charge density.

Tertiary Structure of PreSl(21-47) of Hepatitis B Virus Studied by NMR Spectroscopy

  • Kyeunghee Yu;Cho, Eun-Wie;Shin, Song-Yub;Kim, Kol-Lyong;Kim, Yangmee
    • Journal of the Korean Magnetic Resonance Society
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    • v.4 no.1
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    • pp.41-49
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    • 2000
  • To design more efficient peptide antagonist against the HBV, preSl(21-47) which carries the HBV receptor binding site for hepatocytes was synthesized and the solution structure of preSl(21-47) was investigated using CD spectroscopy and NMR spectroscopy in membrane-mimicking environments. According to CD spectra, preSl(21-47) has a random structure in aqueous solution, while conformational change was induced by addition of TFE and SDS micelle. Tertiary structures as determined by NMR spectroscopy shows that preSl(21-47) has a very flexible structure even in SDS micelle.

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Interaction of Hornet Venom and its Derivatives in the Phospholipid Membrane Environment (말벌 독성 물질과 그 유도체의 인지질 막 환경에서의 상호작용)

  • 이봉헌;박홍재
    • Journal of Environmental Science International
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    • v.7 no.1
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    • pp.62-66
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    • 1998
  • Toxic Mastoparan B(MP-B) which is purified from the venom of the hornet Vespa basalis is a cationic amphlphilic tetradecapeptide. MP-B and Its Ala-substituted analogues were synthesized by solld phase method and the toxic peptide-membrane interactions were examined by circular dichroism(CD) spectra, fluorescence spectra, and leakage abilities in phospholipid membranes. In the presence of phospholipid vesicles, synthetic MP-B and its analogues formed amphiphilic -helical structures, but in the buffer soletion, those exhibited random coil conformation as measured by CD. Fluorescence spectra of MP-B and its analogues which indicated the binding affinity of peptide on phospholipid vesicles showed that the replacement of Lys at position 2 and 11 with Ala caused a remarkable effect in the blue shalt and that at position 2, in the leakage ability of the peptide.

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Production of Peptides Enhancing Calcium Solubility in the Presence of Phosphate Ions In Vitro (In Vitro 상에서인 이온 존재 하에서의 칼슘 용해도를 증대시키는 펩타이드의 생산)

  • 이윤동;이현수
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.485-490
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    • 1997
  • Gluten peptide was produced from corn gluten by enzymatic hydrolysis. This peptide had an ability to increased the solubility of calcium owing to protect calcium ions from forming precipitates of calcium phosphate in the presence of phosphate ions. The solubility of calcium was increased 5.2 times in the presence of 8.3 mg peptide produced by the treatment of papain. These peptides contained high acidic amino acids and fractionated by Delta pack column into fractions No. 1, No. 2 and No. 3. Among them the fraction No. 3 had the highest calcium binding capacity.

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