• Title/Summary/Keyword: Gal2

Search Result 613, Processing Time 0.022 seconds

Purification and Characterization of β-N-Acetylhexosaminidase from Rice Seeds

  • Jin, Yu-Lan;Jo, Yu-Young;Kim, Kil-Yong;Shim, Jae-Han;Kim, Yong-Woong;Park, Ro-Dong
    • BMB Reports
    • /
    • v.35 no.3
    • /
    • pp.313-319
    • /
    • 2002
  • N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

Immunohistochemical studies on the relationship between pineal body and superior cervical ganglia of the Korean native goat (한국재래산양 송과체와 앞쪽목신경절의 관계규명을 위한 면역조직화학적 연구)

  • Lee, Heungshik S.;Lee, In-Se;Song, Seung-hoon;Yoon, Sung-tae;Hwang, In-koo;Lee, Choong-hyun
    • Korean Journal of Veterinary Research
    • /
    • v.40 no.2
    • /
    • pp.197-211
    • /
    • 2000
  • The pineal body have been known to be affected by superior cervical ganglia, and most of its nerve fibers containing peptidergic neurotransmitters have been considered to be originated from this ganglia. To confirm this relationships, some peptidergic neurotransmitters were identified in both of pineal body and superior cervical ganglia of the Korean native goat, which were divided into two group; breeding season and non-breeding season. The localizations of two catecholamine-synthesizing enzymes; tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), were investigated by immunohistochemistry in the superior cervical ganglia and the pineal body of adult Korean native goats. Substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin (GAL) were also identified in these organs by immunohistochemical and double immunofluorescent methods. In superior cervical ganglia, immunoreactivities for TH and DBH were confirmed in the same ganglion cells. The immunoreactivites for SP, VIP(only in male), NPY and GAL were identified in both of ganglion cell bodies and nerve fibers in the ganglia. CGRP immunoreactivity, however, was observed only in nerve fibers. Most NPY- and VIP-immunoreactive(IR) ganglion cells also contained TH. SP and TH were colocalized in the cell bodies, but not in the nerve fibers. TH immunoreactivity was shown in almost all of ganglion cells in the superior cervical ganglia. The immunoreactivity for NPY had some seasonal variation and was stronger in breeding season than in non-breeding season. In pineal body, lots of TH-IR fibers were observed throughout the parenchyma including the pineal stalk and most of them also contained DBH. SP- and NPY-IR fibers were also immunostained with TH or DBH. But a few SP- and NPY-IR fibers were not colocalized with TH or DBH. Exceptionally, a bipolar neuron-like cell was observed to be immunostained with NPY in the pineal body. A few CGRP and GAL-IR fibers were observed, while VIP-IR fibers were not present. It is concluded that most TH- and DBH-IR fibers as well as the peptidergic immunoreactive fibers of the pineal body might be originated from the superior cervical ganglia. Some peptidergic immunoreactive fibers, however, might be come from other regions of brain. We also suggest that NPY in pineal body plays a important role for pineal function. The seasonal variation of NPY immunoreactivity indicates that the synthesis and use of NPY may be different between in breeding and non-breeding seasons.

  • PDF

Zygotic Expression of c-myc Gene in Mouse Early Embryos: Functional Role of c-myc Promoter (생쥐 초기배아에서 c-myc Proto-Oncogene Promoter의 기능적 활성화)

  • Park, Ki-Soo;Kang, Hae-Mook;Shim, Chan-seob;Sun, Woong;Kim, Jae-man;Lee, Young-Ki;Kim, Kyung-jin
    • The Korean Journal of Zoology
    • /
    • v.38 no.4
    • /
    • pp.550-556
    • /
    • 1995
  • The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

  • PDF

Synthesis and Structure of a Second Polymorph of Strontium Germanium Nitride: β-Sr2GeN2

  • Park, Dong-Gon;Gal, Zoltan A.;DiSalvo, Francis J.
    • Bulletin of the Korean Chemical Society
    • /
    • v.26 no.5
    • /
    • pp.786-790
    • /
    • 2005
  • A new ternary strontium germanium nitride, ${\beta}-Sr_2GeN_2$, was obtained as single crystal from constituent elements in molten Na. It crystallizes in space group Cmca (No. 64) with a = 5.441(1) $\AA$, b = 11.377(2) $\AA$, c = 12.229(2) $\AA$, and Z = 8. Its crystal structure is closely related to that of polymorphic companion, ${\alpha}-Sr_2GeN_2$, both of which contain isolated bent anions of ${GeN_2}^{4-}$.

Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells

  • Kim, Seong-Wan;Kim, Seong-Ryul;Park, Seung Won;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.34 no.1
    • /
    • pp.1-5
    • /
    • 2017
  • Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

Synthesis and Properties of Poly(2-ethynyl-N-propargylpyridinium bromide)

  • Gal, Yong Sun;Lee, Won Cheol;Gwi, Tae Rong;Lee, Sang Seop;Bae, Jang Sun;Kim, Bong Sik;Jang, Sang Hui;Jin, Seong Ho
    • Bulletin of the Korean Chemical Society
    • /
    • v.22 no.2
    • /
    • pp.183-188
    • /
    • 2001
  • Ionic conjugated polymer, poly(2-ethynyl-N-propargylpyridinium bromide), was prepared by the cyclopolymerization of 2-ethynyl-N-propargylpyridinium bromide on using various transition metal catalysts, or by thermal methods without adding catalyst. The polymerization of 2-ethynyl-N-propargylpyridinium bromide catalyzed by PdCl2 gave the resulting polymers in relatively high yields. The polymer structure was characterized by various instrumental methods to confirm the conjugated polymer backbone structure carrying cumulated pyridine moiety. The polymers prepared by PdCl2 in DMSO or m-cresol were completely soluble in DMF, DMSO, and formic acid. The inherent viscosities of the resulting polymers were in the range of 0.07-0.19 dL/g. Thermal properties of the polymers were also discussed.

Cloning and expression of human $\beta$$_2$-adrenergic receptor in Saccharomyces cerevisiae

  • 장원진;안진현;고광호;강현삼
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1994.04a
    • /
    • pp.295-295
    • /
    • 1994
  • The human ${\beta}$$_2$-adrenergic receptor (h${\beta}$$_2$AR) contains seven clusters of hydrophobic amino acids suggestive of membrane-spanning domains and its gene is intronless. The genomic gene encoding h${\beta}$$_2$AR has been isolated by polymerase chain reaction. To express h${\beta}$$_2$AR in Saccharomyces cerevisiae, a modified h${\beta}$$_2$AR gene was fused to signal peptide sequence of Killer toxin gene from Kluyveromyces lactics. This fusion gene was expressed under the galactose-inducible GAL10 promoter. The ligand binding experiments showed that the functional h${\beta}$$_2$AR was expressed at a concentration three times as much as that found in Hamster lung.

  • PDF

Synthesis of 2 -Azidoethyl Trisaccharide,$\alpha-D-Gal-(1\righarrow2)-6d-\alpha-D- Altro-Hepp-(1\rightarrow3)-\beta$-D-GlcNAc, an O-Antigenic Repeating Unit of C.jejuni O:23 and O:36

  • Yun, Mi-Kyung;Yoon, Shin-Sook;Shin, Young-Sook;Chun, Keun-Ho;Namshin, Jeong. E
    • Archives of Pharmacal Research
    • /
    • v.27 no.2
    • /
    • pp.143-150
    • /
    • 2004
  • A trisaccharide, the O-antigenic repeating unit of C. jejuni serotype O:23 and O:36, was synthesized as a 2 -azidoethyl glycoside by block addition of perbenzylated thiogalactoside donors to $\alpha$-altroHepp-(1$\rightarrow$3)-GlcNPhth disaccharide acceptor in presence of IDCP promoter. The $\alpha$-linked altroheptopyranoside moiety in the glycosyl acceptor was effectively prepared by Swern oxidation of $\alpha$-mannohepp-(1$\rightarrow$3)-GlcNPhth disaccharide followed by mild reduction with1 $NaCNBH_3$.

Protective Effect of ACTIValoe N-931 Complex, a Mixture of Aloe vera and Silybum marianum, on Experimental Acute Liver Injury

  • Moon, Young-Joo;Cheon, Ho-Jun;Lee, Woo-Cheol;Kim, Hyo-Yeon;Oh, Sun-Tack;Shin, Eun-Ju;Shim, Kyu-Suk;Lee, Sun-Mee
    • Biomolecules & Therapeutics
    • /
    • v.16 no.3
    • /
    • pp.203-209
    • /
    • 2008
  • The aim of this study was to investigate the hepatoprotective effect of $ACTIValoe^{(R)}$ N-931 complex, a mixture of Aloe vera and Silybum marianum, against acute liver injuries. Acute liver damages were induced by intraperitoneal injection of galactosamine (GalN, 700 mg/kg), naphthylisothiocyanate (ANIT, 40 mg/kg) and ethionine (500 mg/kg). $ACTIValoe^{(R)}$ N-931 (85, 170 and 340) was administered orally 48 h, 24 h, 2 h before and 6 h after the injection of hepatotoxins. At 24 h after GalN treatment the levels of serum aminotransferases and hepatic lipid peroxidation were significantly elevated, whereas hepatic glutathione, serum triglyceride (TG) and total cholesterol were decreased. These changes were attenuated by $ACTIValoe^{(R)}$ N-931 complex. The serum aminotransferase activities and total bilirubin significantly increased at 48 h after ANIT treatment, but were attenuated by $ACTIValoe^{(R)}$ N-931 complex. The bile flow was lower after ANIT treatment, which was restored by $ACTIValoe^{(R)}$ N-931 complex. $ACTIValoe^{(R)}$ N-931 complex reduced the ethionine-induced elevated hepatic TG contents. Histopathological analysis revealed that signs of liver injury were prominent at 24 h as result of ethionine injection, demonstrated by extensive areas of fatty change and microvesicular steatosis were observed around cells. These changes were attenuated by $ACTIValoe^{(R)}$ N-931 complex. Our results suggest that the $ACTIValoe^{(R)}$ N-931 complex has a protective effect on acute liver injury.

The Processing of Seasoned and Fermented Oyster and Its Qulaity Changes during the Fermentation (굴 조미 젓갈 제품의 숙성 중 품질 변화에 관한 연구)

  • Lee, Heon-Ok;Rhee, Seong-Kap;Lee, Seong;Kim, Dong-Soo
    • Applied Biological Chemistry
    • /
    • v.44 no.2
    • /
    • pp.81-87
    • /
    • 2001
  • Oyster jeot-gal were prepared in the form of salt-fermented oyster and oyster in soy sauce tentatively and used for investigation the retarding effect of its fermentation in a vacuum from the physicochemical and microbiological points of view. $_PH$ value decreased slightly but amino-N (AN) and volatile basic nitrogen (VBN) increased inversely during the fermentation periods. AN contents were greater in vacuum fermentation than in non-vacuum, whereas VBN were greater in non-vacuum. Total viable cell counts were similar to trend of gentle decrement after increment to some degree but showed higher in non vacuum than in vacuum. In vacuum product, total amino acid contents increased with the elapse of fermentation days or in time of reduction those were higher than in non-vacuum. On the results of chemical analysis, it showed that fermentation was delayed in vacuum and that vacuum fermentation was effective for the shelf-life extension of jeot-gal.

  • PDF