• Title/Summary/Keyword: $Sec61{\beta}$

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Treatment of Epidermal Growth Factor (EGF) enhances Nuclear Maturation of Porcine Oocytes and Stimulates Expression of ER/Golgi Transport Proteins

  • Hwangbo, Yong;Oh, Hae-In;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.21 no.2
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    • pp.131-138
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    • 2017
  • This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta ($Sec61{\beta}$), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVM I) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVM II). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, $Sec61{\beta}$, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVM I or IVM II stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, $Sec61{\beta}$ and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but $Sec61{\beta}$ and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, $Sec61{\beta}$, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of $Sec61{\beta}$ and COPG2 could be changed by EGF in the porcine oocytes during maturation.

사람의 $\beta$$_2$-adrenergic receptor 유전자의 cloning 및 효모에서의 발현

  • 강현삼;고광호
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.61-61
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    • 1993
  • 사람의 혈액의 백혈구로부터 Polymerase Chain Reaction 방법으로 $\beta$$_2$-adrenergic receptor 유전자를 증폭하였다. 이 증폭된 유전자를 pbluescript KS(+)에 cloning하였으며, 제한효소지도 작성과 부분적인 염기배열 확인으로 증폭된 유전자가 사람의 $\beta$$_2$-adrenergic receptor 유전자임을 확인하였다. $\beta$$_2$-adrenergic receptor 유전자를 N말단의 아미노산이 21개 제거되도록 결손시킨후 yeast의 pSec5의 Killer toxin signal sequence의 뒤에 in-frame으로 연결하였다. 현재 효모에서의 발현양상 및 발현된 단백질의 활성을 조사중이다.

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Molecular Cloning of the Sec61p ${\gamma}$ Subunit Homologue Gene from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Eun-Sun;Lee, Heui-Sam;Ahn, Mi-Young;Sohn, Hung-Dae;Ryu, Kang-Sun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.5 no.1
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    • pp.73-77
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    • 2002
  • The Sec61 trimeric complex ($\alpha$,$\beta$, and ${\gamma}$ subunits) is one of the Sec-complex responsible for post-translational protein translocation across the endoplasmic reticulum membrane in diverse organisms. In this study, a cDNA encoding the Sec61p ${\gamma}$ subunit homologue was isolated from the cDNA library of the mole cricket, Gryllotalpa orientalis. Sequence analysis of a 442-bp cDNA clone showed it to contain an open reading frame of 68 amino acid residues consisted of 204-bp. The homologues of the gene were found in the GenBank database in a diverse organism including insect, mammals, fungi, and plants. The deduced amino acid sequence of Sec61p ${\gamma}$ subunit homologue of the mole cricket showed the highest homology to the gene of the singly known insect, Drosophila melanogester (93% identity), and the least homology to that of the baker's yeast, Saccharomyces cerevisiae (37.2%). Phylogenetic analysis also confirmed a close relationship between the insect Sec61p ${\gamma}$ subunit homologues of G. orientalis and D. melanogester. Hydropathy analysis of the cricket mole and published other data suggested that the hydrophobic segment close to C-terminus is predicted to be the putative membrane anchor, Multiple alignment of the Sec61p ${\gamma}$ subunit homologue among several organisms showed the presence of several conserved domains including the conserved proline at position 28.

Role of Golgi Apparatus on Regulation of Sec61β, COPG2 and Epidermal Growth Factor during Oocyte Maturation

  • Oh, Hae-In;Lee, Sang-Hee;Lee, Seunghyung;Lee, Seung Tae;Lee, Eunsong;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.37-41
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    • 2015
  • The oocyte undergoes various events during maturation and requires many substances for the maturation process. Various intracellular organelles are also involved in maturation of the oocyte. During the process glucose is essential for nuclear and cytoplasmic maturation, and adenosine triphosphate is needed for reorganization of the organelles and cytoskeleton. If mitochondrial function is lost, several developmental defects in meiotic chromosome segregation and maturation cause fertilization failure. The endoplasmic reticulum, a store for $Ca^{2+}$, releases $Ca^{2+}$ into the cytoplasm in response to various cellular signaling molecules. This event stimulates secretion of hormones, growth factors and antioxidants in oocyte during maturation. Also, oocyte nuclear maturation is stimulated by growth factors such as epidermal growth factor. This review summarizes roles of organelles with focus on the Golgi apparatus during maturation in oocyte.

Enhanced Nasal Absorption of Ketoconazole by Inclusion with Cyclodextrin (시클로덱스트린과의 포접에 의한 케토코나졸의 비점막 흡수증가)

  • Park, Gee-Bae;Seo, Bo-Youn;Ann, Hong-Jik;Rho, Hyun-Goo;Onn, Yun-Sung;Lee, Kwang-Pyo
    • Journal of Pharmaceutical Investigation
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    • v.24 no.2
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    • pp.95-104
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    • 1994
  • Inclusion complexes of ketoconazole(KT) with ${\alpha}^_$, ${\beta}^_$cyclodextrin(CD) and $dimethy1-{\beta}-cyclodextrin$ (CD) and $dimethy1-{\beta}-cyclodextrin(DM{\beta}CD)$ as nasal absorption enhancer were prepared in 1: 2 molar ratios by freeze-drying and solvent evaporation methods. In order to compare with the intrinsic absorptivity of KT in the jejunum(J) and the nasal cavity(N), the in situ simultaneous perfusion method was employed. The in situ recirculation study revealed that KT-CD inclusion complexes with the greater stability constant and the faster dissolution rate proportionally increased the absorption of KT in the J and N of rats. The rank order of apparent KT permeability$(P_{app}\;:\;cm/sec\;{\time}\;1O^{-5}{\pm}S.E.)$, corrected by surface area of absorption, was $5.10{\pm}0.3(N,\; KT-DM{\beta}CD)$ )> $4.13{\pm}0.4(N,\;KT-{\beta}-CD)$ )> $3.52{\pm}0.2(N,\;KT-{\alpha}-CD)$ )> $2.76{\pm}0.3(J,\; KT-DM{\beta}CD)$ )> $2.61{\pm}0.5(J,\;KT-{\beta}-CD)$ )> $2.42{\pm}0.4(J,\;KT-{\alpha}-CD)$ at pH 4.0. The in crease in permeability of $KT-DM{\beta}CD$ inclusion complex was 2.6 folds in the J and 4.5 folds in the N when the perfusing solution was changed from the buffer(pH 4.0) to saline. The absorption rate of $KT-DM{\beta}CD$ inclusion complex after nasal administration was more rapid than those of ketoconazole alone and $KT-DM{\beta}CD$ inclusion complex after oral administration to rats. In comparision with an oral administration of ketoconazole suspension in corn oil, the relative bioavailability was calculated 137.3% for the oral and 195.0% for nasal $KT-DM{\beta}CD$ inclusion complex in rats. The present results suggest that $KT-DM{\beta}CD$ inclusion complex may serve as a potential nasal absorption enhancer for the nasal delivery of ketoconazole.

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Study on the Travel and Tractive Characteristics of the Two-Wheel Tractor on the General Slope Land(III)-Tractive Performance of Power Tiller- (동력경운기의 경사지견인 및 주행특성에 관한 연구 (III)-동력경운의 경사지 견인성능-)

  • 송현갑;정창주
    • Journal of Biosystems Engineering
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    • v.3 no.2
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    • pp.35-61
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    • 1978
  • To find out the power tiller's travel and tractive characteristics on the general slope land, the tractive p:nver transmitting system was divided into the internal an,~ external power transmission systems. The performance of power tiller's engine which is the initial unit of internal transmission system was tested. In addition, the mathematical model for the tractive force of driving wheel which is the initial unit of external transmission system, was derived by energy and force balance. An analytical solution of performed for tractive forces was determined by use of the model through the digital computer programme. To justify the reliability of the theoretical value, the draft force was measured by the strain gauge system on the general slope land and compared with theoretical values. The results of the analytical and experimental performance of power tiller on the field may be summarized as follows; (1) The mathematical equation of rolIing resistance was derived as $$Rh=\frac {W_z-AC \[1+ \frac{sl}{K} \(\varrho ^{-\frac{sl}{K}-1\)\] sin\theta_1}} {tan\phi \[1+ \frac{sl}{K} \(\varrho ^{-\frac{sl}{K}-1\)\]+\frac{tan\theta_1}{1}$$ and angle of rolling resistance as $$\theta _1 - tan^1\[ \frac {2T(AcrS_0 - T)+\sqrt (T-AcrS_0)^2(2T)^2-4(T^2-W_2^2r^2)\times (T-AcrS_0)^2 W_z^2r^2S_0^2tan^2\phi} {2(T^2-W_z^2r^2)S_0tan\phi}\] $$and the equation of frft force was derived as$$P=(AC+Rtan\phi)\[1+ \frac{sl}{K} \(\varrho ^{-\frac{sl}{K}-1\)\]cos\phi_1 \ulcorner \frac {W_z \ulcorner{AC\[ [1+ \frac{sl}{K} \(\varrho ^{-\frac{sl}{K}-1\)\]sin\phi_1 {tan\phi[1+ \frac{sl}{K} \(\varrho ^{-\frac{sl}{K}-1\]+ \frac {tan\phi_1} { 1} \ulcorner W_1sin\alpha $$The slip coefficient K in these equations was fitted to approximately 1. 5 on the level lands and 2 on the slope land. (2) The coefficient of rolling resistance Rn was increased with increasing slip percent 5 and did not influenced by the angle of slope land. The angle of rolling resistance Ol was increasing sinkage Z of driving wheel. The value of Ol was found to be within the limits of Ol =2\ulcorner "'16\ulcorner. (3) The vertical weight transfered to power tiller on general slope land can be estim ated by use of th~ derived equation: $$R_pz= \frac {\sum_{i=1}^{4}{W_i}} {l_T} { (l_T-l) cos\alpha cos\beta \ulcorner \bar(h) sin \alpha - W_1 cos\alpha cos\beta$$The vertical transfer weight $R_pz$ was decreased with increasing the angle of slope land. The ratio of weight difference of right and left driving wheel on slop eland,$\lambda= \frac { {W_L_Z} - {W_R_Z}} {W_Z} $, was increased from ,$\lambda$=0 to$\lambda$=0.4 with increasing the angle of side slope land ($\beta = 0^\circ~20^\circ) (4) In case of no draft resistance, the difference between the travelling velocities on the level and the slope land was very small to give 0.5m/sec, in which the travelling velocity on the general slope land was decreased in curvilinear trend as the draft load increased. The decreasing rate of travelling velocity by the increase of side slope angle was less than that by the increase of hill slope angle a, (5) Rate of side slip by the side slope angle was defined as $ S_r=\frac {S_s}{l_s} \times$ 100( %), and the rate of side slip of the low travelling velocity was larger than that of the high travelling velocity. (6) Draft forces of power tiller did not affect by the angular velocity of driving wheel, and maximum draft coefficient occurred at slip percent of S=60% and the maximum draft power efficiency occurred at slip percent of S=30%. The maximum draft coefficient occurred at slip percent of S=60% on the side slope land, and the draft coefficent was nearly constant regardless of the side slope angle on the hill slope land. The maximum draft coefficient occurred at slip perecent of S=65% and it was decreased with increasing hill slope angle $\alpha$. The maximum draft power efficiency occurred at S=30 % on the general slope land. Therefore, it would be reasonable to have the draft operation at slip percent of S=30% on the general slope land. (7) The portions of the power supplied by the engine of the power tiller which were used as the source of draft power were 46.7% on the concrete road, 26.7% on the level land, and 13~20%; on the general slope land ($\alpha = O~ 15^\circ ,\beta = 0 ~ 10^\circ$) , respectively. Therefore, it may be desirable to develope the new mechanism of the external pO'wer transmitting system for the general slope land to improved its performance.l slope land to improved its performance.

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