• The Korean Journal of Physiology
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• 제23권1호
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• pp.99-108
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• 1989

The effects of prostaglandin $(PGF_{2{\alpha}})$ on the contractility of vascular smooth muscle were investigated in the helical strip of the rabbit aorta. The aortic strip was immersed in the phosphate-buffered Tyrode's solution which was equilibrated with 100% $O_{2}$ at $35^{\circ}C$ and its isometric tension was measured. The contraction was induced by $(PGF_{2{\alpha}})$, norepinephrine (NE), or potassium (40 mM) in the nomal Tyrode's solution (1 mM, $Ca^{2+}$) or $Ca^{2+}-free$ Tyrode's solution. Effects of verapamil and phentolamine on the contraction were also observed. The aortic strip began to contract at the concentration of $5\;{\mu}g%$ and reached the maximal contraction at the concentration of $150\;{\mu}g%$ $(PGF_{2{\alpha}})$. The maximal contraction was corresponded respectively to $52.2{\pm}3.0%$ and $81.5{\pm}3.5%$ of maximal contraction by NE $(1{\times}10^{-5}M)$ and 40 mM $K^{+}$. And the maximal contractions by $(PGF_{2{\alpha}})$ or NE were induced at the concentration of about 1 mM $Ca^{2+}$. $(PGF_{2{\alpha}})$ induced the contraction of aortic strip even after induction of contraction by 40 mM $K^{+}$ and the contraction by $(PGF_{2{\alpha}})$ was not blocked by the ${\alpha}-receptor$ blocker, phentolamine. And the contraction by the $(PGF_{2{\alpha}})$ was inhibited partially by a verapamil at the concentration of $1{\times}10^{-5}M$ and the contraction began to increase at the concentration of $1{\times}10^{-4}M$ verapamil. Whereas the contraction by NE was completely blocked by verapamil. Though both the $(PGF_{2{\alpha}})$ and NE induced the contraction in the $Ca^{2+}-free$ Tyrode's solution, the peak tension was not maintained. But the rate of tension decline was lower in the contraction by $(PGF_{2{\alpha}})$ than in that by NE. The verapamil did not inhibit the contraction by $(PGF_{2{\alpha}})$ in the $Ca^{2+}-free$ Tyrode's solution and increased the contraction at the concentration of above $1{\times}10^{-4}M$. The NE-induced contraction in the $Ca^{2+}-free$ Tyrode's solution was inhibited completely by a verapamil. From the above results it is suggested that the contraction induced by $(PGF_{2{\alpha}})$ results from the promotion of the both $Ca^{2+}$ influx and the intracellular $Ca^{2+}$ release by different way from NE.

• 한국자기공명학회논문지
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• 제7권1호
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• pp.37-45
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• 2003

Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by mic2 gene. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 Type II, we have initiated studies on hCD99$\^$TMcytoI/ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$\^$TMytoI/ in the presence of 200mM DPC and CHAPS displayed an existence ${\alpha}$-helical conformation, showing that it could form an ${\alpha}$-helix under membrane environments. In addition, we have found that the cytoplasmic domain of CD99 would form symmetric dimmer in the presence of transmembrane domain. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$\^$TMcytoI/, the dimerization or oligomerization would play an important role in its biological function.

• 한국자기공명학회논문지
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• 제11권2호
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• pp.73-84
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• 2007

MTH1821 (UniProtKB/TrEMBL ID O27849) is a 96-residue hypothetical protein from the open reading frame of Methanobacterium thermoautotrophicum H one of the target organisms of structural genomics pilot project. Proteins which contain conserved sequence compared with MTH1821 have not been discovered yet and the functional and structural information for MTH1821 is not available. Here, we present the sequence-specific backbone resonance using multidimensional heteronuc1ear NMR spectroscopy and propose the secondary structure using GetSBY software. The backbone resonances of N, HN, $C_{\alpha}$, $C_{\beta}$, CO and $H_{\alpha}$ which are necessary for a prediction of secondary structure by GetSBY were assigned about 98% (557/568). The secondary structure of MTH1821 confirmed that it is comprised of four strand regions and two helical regions. This report will provide a valuable resource for the calculation solution structure of MTH1821 and for the other hypothetical protein that is targeted for structural-based functional discovery.

• 한국자기공명학회논문지
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• 제16권1호
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• pp.22-33
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• 2012

Tid1, belonging to the Hsp40/DnaJ family of proteins, functions as a cochaperone of cytosolic and mitochondrial Hsp70 proteins. In particular, the N-terminal J-domain of Tid1 (Tid1-JD) constitutes the major binding sites for proteinprotein interactions with client proteins, including p53, as well as its partner chaperone, Hsp70. In the present study, soluble, recombinant protein of Tid1-JD could be obtained by using the pCold vector system, and backbone NMR assignments were completed using the isotope $[^{13}C/^{15}N]$-enriched protein. Far-UV CD result implied that Tid1-JD is an ${\alpha}$-helical protein and the secondary structure determined using chemical shift data sets indentified four ${\alpha}$-helices with a loop region containing the HPD (conserved tripeptide of His, Pro and Asp) motif. Additionally, NMR spectra under different conditions implied that the HPD motif, which is a critical region for protein-protein interactions of Tid1-JD, would possess dynamic properties.

• Bulletin of the Korean Chemical Society
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• 제35권12호
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• pp.3627-3631
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• 2014

In this study we examined two ester-containing cross-links, hex-2-enyl acetate and hex-2-enyl propionate, as new cross-linking systems for helix stabilization of short peptides. We demonstrated that these hexenyl ester cross-links can be readily installed via a ruthenium-mediated ring-closing metathesis reaction of L-aspartic acid 4-allyl ester or L-glutamic acid 5-allyl ester at position i and (S)-2-(4'-pentenyl)alanine at position i+4 using second generation Hoveyda-Grubbs catalyst at $60^{\circ}C$. Between these two cross-links, we found that the hex-2-enyl propionate significantly stabilizes the ${\alpha}$-helical conformations of short model peptides. The helix-stabilizing effects of the hex-2-enyl propionate tether appear to be as powerful as Verdine's i,i+4 all-hydrocarbon stapling system, which is one of the most widely used and the most potent helix-stabilizing cross-linking systems. Furthermore, the hex-2-enyl propionate bridge is reasonably robust against non-enzymatic hydrolytic cleavage at a physiological pH. While extended studies for probing its chemical scopes and biological applications are needed, we believe that this new helix-stabilizing system could serve as a useful chemical tool for understanding protein folding and designing conformationally-constrained peptide drugs.

• 대한의용생체공학회:의공학회지
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• 제17권3호
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• pp.297-304
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• 1996

Although collagen is still considered to be a poor immunogen, animals can produce antibodies to a number of different sites in the collagen molecule. In type I collagen, three classes of antigenic determinants have been described those are recogrlized as different degrees in different species. These are essentially composed of helical, conformation-dependent antigenic determinants and terminal, nonhelical antigenic determinants, and finally central antigenic determinants exposed only after denaturation of the collagen molecule. To utilize collagen as implantable biomateriall human e61bryonic collagen, ten immunological to body, was purified from human umbilical cords and found to contain [$\alpha$1(I)]$_2$. [$\alpha$2(I). Each step of purification were observed by polarized light microscope and analyzed through SDS-PAGE. The conclusious are follows; 1 . The purified collagen revealed gradual fiber indenties on each step of purification by polarized microscope. 2. The structual changes of extracted collagen as removed telopeptide were confirmed by SDS-PAGE.

• Bulletin of the Korean Chemical Society
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• 제24권10호
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• pp.1478-1484
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• 2003

CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.

• 한국미생물·생명공학회지
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• 제31권4호
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• pp.311-321
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• 2003

세균은 지방을 분해할 수 있는 다양한 리파제를 생산한다. 리파제는 반응조건에 따라서 지방의 합성도 수행할 수 있는데 , 이러한 효소반응과정에서 고도의 기질특이성과 위치특이성 및 입체특이성을 보이기 때문에 제약산업과 정밀화학산업에서 효소촉매로서 널리 사용되고 있다. 지금가지 200종류 이상의 리파제효소가 보고되었으며, 이것들은 효소생산기원과 아미노산 상동성을 기준으로 6개의 family로 분류된다. 지난 10년 간 세균 리파제 6종에 대한 3D구조가 밝혀졌다. 이것들은 모두 중심부분에$\alpha/\beta$폴딩구조와 세린, 히스티딘, 아스팔틴산으로 구성된 활성부위를 공통적으로 갖고 있다. 활성부위를 양친성 $\alpha$나선구조가 뚜껑처럼 덮고 있으며, 물과 오일의 경계면을 만나면, 이 뚜껑이 열리고 효소활성이 크게 증가하는 '계면활성화' 현상을 보인다. P. cepacia 리파제 구조에는 기질과 결합하는 4개의 포켓이 있는데 이중 하나는 옥시음이온 구멍이고, 다른 세 개는 기질의 sn-1, sn-2, sn-3 지방산과 결합하는 부위이다. 이 포켓의 크기와 방향 및 소수성정도에 의해서 효소의 기질특이성과 입체특이성이 결정된다. 현재 이러한 구조연구를 기반으로 사용목적에 따른 맞춤 효소를 생산하기 위한 효소 개량연구가 활발히 진행되고 있다.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.3928-3932
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• 2011

We synthesized peptide-resin conjugates (1 and 2) by immobilizing ${\beta}$-sheet antibacterial peptide and ${\alpha}$ helical antibacterial peptide on PEG-PS resin, respectively. Conjugate 1 showed considerable antibacterial activity in various conditions, whereas conjugate 2 did not exhibit antibacterial activity. The growths of various bacteria were inhibited by conjugate 1 even at lower concentrations than MIC. Conjugate 1 killed bacteria at MIC and had a potent synergistic effect with current antibacterial agents such as vancomycin and tetracycline, respectively. Overall results indicate that polymer surface modification using antibacterial ${\beta}$ sheet peptide is a powerful way to prevent microbial contamination on polymer surfaces.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.4021-4026
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• 2011

Functions of the luteinizing hormone releasing hormone (LHRH) and its induced release by divalent metal ions have received great attention because this neurotransmitter subsequently regulates the secretion of luteinizing hormone (LH). Metal-LHRH complexes were synthesized by addition of various Cu(II),Ni(II),Zn(II) ions into LHRH in order to understand how the induced release of LHRH is possible. The degree of complexation was monitored by $^1H$, $^{13}C$-NMR chemical shifts, and final products were identified by Mass spectrometry. Solutionstate structure determination of Zn(II)-LHRH out of metal-complexes was accomplished by using NMR and NMR-based distance geometry (DG). Interproton distance information from nuclear Overhauser effect spectroscopy was utilized for structure determination. Structure obtained in this study has a cyclic conformation exhibiting a specific ${\alpha}$-helical turn with residue numbers His[2]-Leu[7] out of 10 amino acids. Comparison of chemical shifts and EPR studies of Ni(II),Cu(II)-LHRH complexes exhibit that these metal complexes have 4-coordination geometry.