• Title/Summary/Keyword: Helix 8

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NMR Studies of the Conformation and Stability of the 4' - Aminomethyl - 4,5',8 - Trimethylpsoralen (AMT) Cross - Linked DNA Octamer Duplex, $d(GGGTACCC)_2$

  • Lee, Joon-Hwa;Hwang, Geum-Sook;Choi, Byong-Seok
    • BMB Reports
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    • v.30 no.6
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    • pp.421-425
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    • 1997
  • The 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT) has been used as intercalating DNA binding drugs in the photo-chemotherapy of skin diseases. The conformation and stability of DNA octamer duplex, $d(GGGTACCC)_2$, cross-linked with AMT has been studied by NMR spectroscopy. All the proton resonances of the psoralen cross-linked octamer were assigned and meting temperature studies were carried out based on the assignment of the proton resonances. The aromatic proton chemical shift data suggest that the conformation of the helix cross-linked with psoralen is destabilized more to furanside of the psoralen. possibly due to the protrusion of the aminomethyl side chain of the psoralen.

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Online Social Support: Which Posts Were Answered?

  • Chang, Hui-Jung
    • Journal of Contemporary Eastern Asia
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    • v.8 no.1
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    • pp.31-46
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    • 2009
  • The purpose of the study was to find out which posts were answered in a text-based computer-mediated social support group. Specifically, the present study examined the effects of two variables on support-seeking behaviors: support-seeking strategies and gender. A revised typology of support-seeking strategies originally proposed in the Sensitive Interaction Systems Theory (SIST) model was employed for the study. Data were collected from the PTT psychosis discussion group, the largest BBS in the Chinese-speaking community, for a period of 30 months from February 2004 to July 2006. In general, the results indicated that posts with more asking, less crying and less hinting were answered more than posts with more hinting, more crying and less asking. However, although different support-seeking strategies did affect support-seeking behaviors, gender did not have an impact on which posts were answered.

In Vitro Evolution of Lipase B from Candida antarctica Using Surface Display in Hansenula polymorpha

  • Kim, So-Young;Sohn, Jung-Hoon;Pyun, Yu-Ryang;Yang, In-Seok;Kim, Kyung-Hyun;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1308-1315
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    • 2007
  • Lipase B from Candida antarctica (CalB) displayed on the cell surface of H. polymorpha has been functionally improved for catalytic activity by molecular evolution. CalB was displayed on the cell surface by fusing to a cell-wall anchor motif (CwpF). A library of CalB mutants was constructed by in vivo recombination in H. polymorpha. Several mutants with increased whole-cell CalB activity were acquired from screening seven thousand transformants. The two independent mutants CalB 10 and CalB 14 showed an approximately 5 times greater whole-cell activity than the wild-type. When these mutants were made as a soluble form, CalB 10 showed 6 times greater activity and CalB 14 showed an 11 times greater activity compared with the wild-type. Sequence analyses of mutant CALB genes revealed amino acid substitutions of $Leu^{278}Pro$ in CalB10 and $Leu^{278}Pro/Leu^{219}Gln$ in CalB14. The substituted $Pro^{278}$ in both mutants was located near the proline site of the ${\alpha}$10 helix. This mutation was assumed to induce a conformational change in the ${\alpha}$10 helix and increased the $k_{cat}$ value of mutant CalB approximately 6 times. Site-directed mutagenized CalB, LQ ($Leu^{219}Gln$) was secreted into the culture supernatant at an amount of approximately 3 times more without an increase in the CalB transcript level, compared with the wild-type.

Binding Specificity of Philyra pisum Lectin to Pathogen-Associated Molecular Patterns, and Its Secondary Structure

  • Park, Byung Tae;Kim, Byung Sun;Park, Heajin;Jeong, Jaehoon;Hyun, Hanbit;Hwang, Hye Seong;Kim, Ha Hyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.6
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    • pp.547-551
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    • 2013
  • We recently reported a Philyra pisum lectin (PPL) that exerts mitogenic effects on human lymphocytes, and its molecular characterization. The present study provides a more detailed characterization of PPL based on the results from a monosaccharide analysis indicating that PPL is a glycoprotein, and circular dichroism spectra revealing its estimated ${\alpha}$-helix, ${\beta}$-sheet, ${\beta}$-turn, and random coil contents to be 14.0%, 39.6%, 15.8%, and 30.6%, respectively. These contents are quite similar to those of deglycosylated PPL, indicating that glycans do not affect its intact structure. The binding properties to different pathogen-associated molecular patterns were investigated with hemagglutination inhibition assays using lipoteichoic acid from Gram-positive bacteria, lipopolysaccharide from Gram-negative bacteria, and both mannan and ${\beta}$-1,3-glucan from fungi. PPL binds to lipoteichoic acids and mannan, but not to lipopolysaccharides or ${\beta}$-1,3-glucan. PPL exerted no significant antiproliferative effects against human breast or bladder cancer cells. These results indicate that PPL is a glycoprotein with a lipoteichoic acid or mannan-binding specificity and which contains low and high proportions of ${\alpha}$-helix and ${\beta}$-structures, respectively. These properties are inherent to the innate immune system of P. pisum and indicate that PPL could be involved in signal transmission into Gram-positive bacteria or fungi.

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility

  • Bai, Mingmei;Qin, Guixin;Sun, Zewei;Long, Guohui
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1159-1165
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    • 2016
  • The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as ${\alpha}$-helices and ${\beta}$-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility ($p{\leq}0.003$); moreover, the relatively quantitative amounts of ${\alpha}$-helices, random coils, and ${\alpha}$-helix to ${\beta}$-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility ($p{\leq}0.004$). On the other hand, the percentage of ${\beta}$-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001) and solubility (p = 0.002). These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the ${\alpha}$-helix-to-${\beta}$-sheet ratio can be used to predict the nutritional value of feed proteins.

Correction of Cryptotia by Triangular V-Y Advancement Flap and Rhomboid Flap (삼각형 V-Y피판 및 장사방형피판을 이용한 매몰귀의 교정)

  • Lee, Joon-Moon;Seo, Dong-Lin;Dhong, Eun-Sang;Yoon, Eul-Sik
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.639-643
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    • 2010
  • Purpose: Cryptotia is a congenital deformity in which the upper third of the auricle is buried under the temporal skin. It is rare in Caucasians, yet it is more common in Asians. Although a variety of methods to treat cryptotia have been introduced, there is still no acceptable single procedure that can successfully manage this deformity in its entity. We present a triangular V-Y advancement flap and rhomboid flap for correcting cryptotia that can overcome the diverse shortcomings of the conventional methods. Methods: This operative method was used to correct 18 auricles in patients ranging in age from 4 to 33 years. A triangular flap was prepared over the auricle by making a skin incision according to Ohmori's method. Then a rhomboid flap with a side length of about 8 to 10 mm that sets the lower portion as a pedicle in the anterior region was prepared to supplement the contracted portion of the helix. The cartilage deformity was corrected by the banner flap or the radiating cartilage incisions with cartilage graft or high density polyethylene graft. Results: We have treated 16 patients with severe cryptotia using this method and have obtained good aesthetic results. All cases showed widened scaphoid fossa and smooth triangular fossa of antihelix. There were no major postoperative complications, such as necrosis or infection of the flaps. Conclusion: Correction of cryptotia using the triangular V-Y advancement flap and rhomboid flap is useful a method for certain conditions, when a severe contraction of the helix is present.

Antitumor Toxic Protein Abrin and Abrus Agglutinin

  • Liu, Chao-Lin;Lin, Jung-Yaw
    • Toxicological Research
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    • v.17
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    • pp.109-115
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    • 2001
  • Abrus agglutinin was purified from the kernels of Abrus precatorius by Sepharose 4B affinity column chromatography followed by Sephadex G-100 gel filtration column chromatography. About 1.25 g of abrus agglutinin was obtained from 1 kg of the kernels. The LD$_{50}$ of abrus agglutinin is 5 mg/kg of body weight, which is less toxic than that of abrin, 20$\mu\textrm{g}$/kg body weight. The amino acid sequence of abrus agglutinin was determined by protein sequencing techniques and deduced from the nucleotide sequence of a cDNA clone encoding full length of abrus agglutinin. There are 258 residues, 2 residues and 267 residues in the A-chain, the linker peptide and the B-chain of abrus agglutinin, respectively. Abrus agglutinin had high homology to abrin-a (77.8%). The 13 amino acid residues involved in catalytic function, which are highly conserved among abrin and ricin, were also conserved within abrus agglutinin. The protein synthesis inhibitory activity of abrus agglutinin ($IC_{50}$/ = 3.5 nM) was weaker than that of abrin-a (0.05 nM). By molecular modeling followed by site-directed mutagenesis showed that Pro199 of abrus agglutinin A-chain located in amphipathic helix H and corresponding to Asn200 of abrin A-chain, can induce bending of helix H. This bending would presumably affect the binding of abrus agglutinin A-chain to its target sequence GpApGpAp, in the tetraloop structure of 285 r-RNA subunit and this could be one of major factors contributing to the relatively weak protein synthesis inhibitory activity and toxicity of abrus agglutinin.n.

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Purification of Caudal-Related Homeodomain Transcription Factor and Its Binding Characterization

  • Jeong, Mi-Suk;Hwang, Eun-Young;Kim, Hyun-Tae;Yoo, Mi-Ae;Jang, Se-Bok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1557-1564
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    • 2009
  • Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

A Study on the X-ray Diffraction of Rabbit Glycerin Muscle by Spin Labeled on SH (SH에 Spin Label한 Rabbit Glycerin처리근육의 X선 회절에 관한 연구)

  • 김덕술;송주영
    • Journal of Life Science
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    • v.8 no.6
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    • pp.681-686
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    • 1998
  • IASL(iodo acetamide) and MSL(maleimide) disordered the orderly helix arrangement of myosin in the rest state of spin level. Especially the effect of IASL was great. Equatorial reflection(10,11) change inferred that myosin head was moved to the vicinity of actin filament by spin level. The intensity change of 143 $\AA$ and 72 $\AA$ could offer infor-mation of the mass projection of population of myosin heads along the filament axis. The slope of intensity profile of the mass projection of 143 $\AA$ and reflection of IASL is appeared and that of MSL is appeared sharply. The dec-rease of 215 $\AA$ reflection intensity the periodical characteristic of 143 $\AA$ reflection by spin label. The raise of MSL actin reflection at 51 $\AA$ and 59 $\AA$ in the actin reflection change refers that the shifted myosin head binds a certain actin or changes an actin structure by spin label effect. Because iodo acetamide has a tendency to decease the actin reflection, actin dose not bind myosin head. From this result, we could conclude that LASL and MSL are spin labeled on SH of myosin head and disordered the helix arrangement of actin.

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Biochemical Properties and Antibacterial Activity of Lactoferrin from Korean Native Cow (한우 Lactoferrin의 생화학적 특성 및 항균 활성)

  • Yang, Hui-Jin;Lee, Su-Won
    • Journal of Dairy Science and Biotechnology
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    • v.23 no.1
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    • pp.1-8
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    • 2005
  • The purpose of this study was to demonstrate biochemical properties and antibacterial activity of lactoferrin(Lf) obtained from the colostrum of Korean native cow. Lactoferrin was isolated from the colostrum of Korean native cow by purification steps using batch extraction, ion exchange chromatography, gel filtration chromatography, affinity chromatography. Other whey protein components that is similar molecular weight and affinity to lactoferrin were gradually removed from crude Korean Native cow's lactoferrin during the purification steps. The molecular weight of the purified Korean native cow's Lf(K-Lf) was 81 kDa, the isoelectric point was 9, and the content of iron was 0.56mg/g, which is indicated that iron saturation of the K-Lf was 40.6%. Amino acid composition and a-helix content were different K-Lf from bovine Lf(B-Lf). Antibacterial activity of E. coli O111 by K-Lf was lower than that of B-Lf. A minimal inhibitory concentration(MIC) of K-Lf and B-Lf was 2.75mg/ml and 1.5mg/ml respectively.

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