• Title/Summary/Keyword: $H^+/K^+-ATPase$

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Analysis of Differentially Expressed Proteins in Bovine Longissimus Dorsi and Biceps Femoris Muscles

  • Kim, S.M.;Park, M.Y.;Seo, K.S.;Yoon, D.H.;Lee, H.-G.;Choi, Y.J.;Kim, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1496-1502
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    • 2006
  • Skeletal muscle contains slow and fast twitch fibers. These skeletal muscle fibers express type I and type II myosin, respectively, and these myosin isoenzymes have different ATPase activity. The aim of this study was to investigate protein profiles of bovine skeletal muscles by proteomic analysis. Fifty seven spots of distinct proteins were excised and characterized. The expression of sixteen spots was differed in longissimus dorsi muscle with a minimal 2-fold change compared to biceps femoris muscle. The majority of differentially expressed proteins belonged to metabolic regulation-related proteins such as glyceraldehyde 3-phosphate dehydrogenase, triosephosphate isomerase and carbonic anhydrase 3. The real time-PCR assay confirmed an increase or induction of specific genes: RGS12TS isoform, GAPDH, triosephosphate isomerase and carbonic anhydrase. These results suggest that the expression of metabolic proteins is under a specific control system in different bovine skeletal muscle. These observations could have significant implications for understanding the physiological regulation of bovine skeletal muscles.

Effect of PCMB on Organic Ion Transport in Rabbit Renal Cortical Slices (토끼 신피질 절편에서 PCMB가 유기이온의 이동에 미치는 영향)

  • Park, In-Cheol;Kim, Tae-In;Jung, Dong-Keun;Kim, Young-Keun
    • The Korean Journal of Physiology
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    • v.24 no.2
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    • pp.345-352
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    • 1990
  • To determine the role of sulfhydryl group in transport of organic ions across the basolateral membrane of renal proximal tubules, effect of p-chloromercuribenzoic acid (PCMB) on the transport of tetraethylammonium (TEA) and p-aminohippurate (PAH) was studied in rabbit renal cortical slices. PCMB caused irreversible inhibition of TEA and PAH uptake in a dose-dependent manner, with $I_{50}$ value (concentration for 50% inhibition) of $30\;{\mu}M$ for TEA and $75\;{\mu}M$ for PAH. Kinetic analysis of TEA and PAH uptakes showed that PCMB decreased Vmax $(62.35\;vs.\;28.32\;n\;mole/g{\cdot}min\;fur\;TEA:\;385.24\;vs.\;170.36\;n\;mole/g{\cdot}min\;for\;PAH)$ without changing Km. The inhibitory action of PCMB on TEA and PAH uptakes was independent of pH of the pretreatment medium. The inhibitory effect of PCMB on the uptake of TEA or PAH was prevented by dithiothreitol, but not by the substrate. PCMB inhibited Na-K-ATPase activity in a dose-dependent manner with $I_{50}$ value of $50\;{\mu}M$, which is similar to those for TEA and PAH uptake. These results suggest that PCMB inhibits the transport of organic cations and anions in the renal basolateral membrane by directly affecting the SH-group in the transporter molecules or secondly by altering the Na-K-ATPase activity.

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Effects of Inositol 1,4,5-triphosphate on Osteoclast Differentiation in RANKL-induced Osteoclastogenesis

  • Son, A-Ran;Kim, Min-Seuk;Jo, Hae;Byun, Hae-Mi;Shin, Dong-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.1
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    • pp.31-36
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    • 2012
  • The receptor activator of NF-${\kappa}B$ ligand (RANKL) signal is an activator of tumor necrosis factor receptor-associated factor 6 (TRAF6), which leads to the activation of NF-${\kappa}B$ and other signal transduction pathways essential for osteoclastogenesis, such as $Ca^{2+}$ signaling. However, the intracellular levels of inositol 1,4,5-trisphosphate ($IP_3$) and $IP_3$-mediated cellular function of RANKL during osteoclastogenesis are not known. In the present study, we determined the levels of $IP_3$ and evaluated $IP_3$-mediated osteoclast differentiation and osteoclast activity by RANKL treatment of mouse leukemic macrophage cells (RAW 264.7) and mouse bone marrow-derived monocyte/macrophage precursor cells (BMMs). During osteoclastogenesis, the expression levels of $Ca^{2+}$ signaling proteins such as $IP_3$ receptors ($IP_3Rs$), plasma membrane $Ca^{2+}$ ATPase, and sarco/endoplasmic reticulum $Ca^{2+}$ ATPase type2 did not change by RANKL treatment for up to 6 days in both cell types. At 24 h after RANKL treatment, a higher steady-state level of $IP_3$ was observed in RAW264.7 cells transfected with green fluorescent protein (GFP)-tagged pleckstrin homology (PH) domains of phospholipase C (PLC) ${\delta}$, a probe specifically detecting intracellular $IP_3$ levels. In BMMs, the inhibition of PLC with U73122 [a specific inhibitor of phospholipase C (PLC)[ and of $IP_3Rs$ with 2-aminoethoxydiphenyl borate (2APB; a non-specific inhibitor of $IP_3Rs$) inhibited the generation of RANKL-induced multinucleated cells and decreased the bone-resorption rate in dentin slice, respectively. These results suggest that intracellular $IP_3$ levels and the $IP_3$-mediated signaling pathway play an important role in RANKL-induced osteoclastogenesis.

NMR Studies on N-terminal Domain of DNA2

  • Jung, Young-Sang;Lee, Kyoung-Hwa;Jung, Jin-Won;Lee, Weontae
    • Journal of the Korean Magnetic Resonance Society
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    • v.4 no.2
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    • pp.74-81
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    • 2000
  • Saccharomyces cerevisiae Dna2 protein has biochemical activities: DNA-dependent ATPase, DNA helicase and DNA nuclease and is essential for cell viability. Especially, Pro$\^$504/ is determined as an important residue in ATPase, helicase, and nuclease activity. We synthesized and determined the three-dimensional solution structure of N-terminal domain comprising residues of Val$\^$501/ -_Phe$\^$508/ (Dna2$\^$pep/) using two-dimensional $^1$H-NMR and dynamical simulated annealing calculations. On the basis of a total of 44 experimental restraints including NOEs, $^3$J$\_$$\alpha$$\beta$/ and $^3$J$\_$$\alpha$$\beta$/ coupling constants, the solution structures of Dna2$\^$epe/ were calculated with the program CNS. The 23 lowest energy structures were selected out of 50 final simulated-annealing structures. The atomic RMSDs of the final 23 structures fur the individual residues were calculated with respect to the average structure. The mean RMSDs for the 23 structures were 0.042 nm for backbone atoms and 0.316 nm for all heavy atoms, respectively. The Ramachandran plot indicates that the $\Phi$, Ψ angles of the 23 final structures are properly distributed in energetically acceptable regions. Solution structure of Dna2$\^$pep/ showed a single unique turn spanning residues of Asn$\^$503/ Val$\^$506/.

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Effect of Hwangryunhaedok-tang on Blood Pressure and Renal Functions in Spontaneously Hypertensive Rats (황연해독탕(黃連解毒湯)이 자발적(自發的) 고혈압(高血壓) 백서(白鼠)의 혈압(血壓) 신장(腎臟) 기능(機能)에 미치는 영향(影響))

  • Kook Yoon-Bum
    • Herbal Formula Science
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    • v.10 no.1
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    • pp.113-129
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    • 2002
  • The present study designed to investigate whether hwangryunhaedok-tang show an anti-hypertensive effect and elucidate its possible mechanism in spontaneously hypertensive rats. The systolic blood pressures (SBP) were significantly decreased as an oral administration of hwangryunhaedok-tang compared with their control group. The urine volume was significantly increased by administration of hwangryunhaedok-tang but urinary sodium (UNaV), potassium (UKV), chloride excretion (UCIV) were not remarkably affected. The urinary creatinine excretion rate (UcrV) was significantly increased in rats administered with hwangryunhaedok-tang in association with increase of creatinine clearance (Ccr). The urine osmolality (Uosmol) was significantly decreased in SHR administered with hwangryunhaedok-tang without being changed in solute-free water reabsorption ( TcH20). The expressions of Aquaporin 2 (AQP-2). 3 and ${\alpha}\;1$, ${\beta}\;1$ subunits of Na.K-ATPase were determined by Western blot analysis to assess the role of these proteins in association with changes of renal functions in SHR administered with hwangryunhaedok-tang. The expression of AQP-2 and 3 protein was significantly down-regulated in the kidney of SHR administered with hwangryunhaedok-tang compared with those in control rats without being altered expression of ${\alpha}\;1$, ${\beta}\;1$ subunits of Na,K-ATPase. In the in vitro assay, Angiotensin converting enzyme (ACE) was inhibited by hwangryunhaedok-tang in a dose-dependent manner. Berberine and/or palmatine, which are well known as a main components of hwangryunhaedok-tang, also have an ACE inhibitory effects in a dose-dependent manner. Taken together, these results suggest that hwangryunhaedok-tang lowered blood pressure through the increase of diuresis caused by down-regulation of water channels and the inhibition of Angiotensin converting enzyme.

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Effect of Radish on HeLa Cell Vacuolation Induced by Helicobacter pylori Cytotoxin (HeLa세포에서 Helicobacter pylori 독소에 의한 공포형성에 미치는 무의 효과)

  • Shon, Yun-Hee;Surh, Jung-Ill;Chung, Yu-Jin;Park, In-Kyung;Kim, Ho-Chang;Hwang, Cheorl-Weon;Kim, Cheorl-Ho;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.35 no.3 s.138
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    • pp.250-254
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    • 2004
  • Helicobacter pylori (H. pyroli) infection it associated with type B gastritis, peptic uler disease, and gastric cancer. The vacuolation of cells induced by H. pylori is thought to be essential for the initiation and maintenance of gastric infection. The roles of H. pylori cytotoxin, urease, and ammonia in the vacuolation of HeLa cells were determined. H. pylori toxin induced vacuolation of HeLa cells. Korean and Japanese radishes significantly prevented the vacuolation of HeLa cells induced by H. pylori toxin. The urease activity in vacuolated cells was also decreased with Korean and Japanese radishes. H. Pylori toxin-induced vacuolation was inhibited by vacuolar type ATPase inhibitors (bafilomycin and N-ethylmaleimide). However, further investigation is required to determine the mechanisms of radish for the inhibition of vacuole formation of eukaryotic cells in response to the H. pylori toxin.

Succinylation of Myofibrillar Protein of PSE(pale, soft, exudative) Porcine Muscle and it's Functional Characteristics (PSE 돈육으로 부터 추출한 근원섬유단백질의 석시닐화와 그 기능적 특성)

  • Sung, Sam-Kyung
    • Korean Journal of Food Science and Technology
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    • v.16 no.3
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    • pp.353-357
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    • 1984
  • The myofibril prepared from PSE (pale, soft, exudative) porcine meat was modified by reacting with succinic anhydride and the chemical and functional properties of modified myofibrils were investigated. $No\;Ca^{2+}-and\;Mg^{2+}-ATPase$ activity were observed irrespective of the degree of succinylation. Isoelectric point of the succinylated myofibril changed to around pH 3 from the pH 5 of unmodified myofibril. Salt soluble property was not affected by changing the salt concentration. The modified myofibril in aqueous solution did not coagulate during heating at $98^{\circ}C$ for 10 min. Water absorption ability was not improved but emulsion capacity was improved a little by succinylation.

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A Histochemical Study of the Masseter Muscle Fibers of White Rat with Unilateral Edentulous Jaw (편측 무치악이 백서 교근 근섭유에 미치는 영향에 관한 조직화학적 연구)

  • Hong-Ryeol Ryu;Sung-Woo Lee
    • Journal of Oral Medicine and Pain
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    • v.13 no.1
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    • pp.61-69
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    • 1988
  • The study of the muscle fiver composition and the muscle fiver type conversion during unilateral edentulous condition was undertaken in the rostral superficial masseter muscle of the whiter rat. 16 4-week-old male white rats weighting approximately 130gm that crowns of left upper and lower molare were cut intentionally, were divided into 4 groups (one control group and 3 experimental groups). After experimental groups were sacrificed by etherization in 6 days($E_1$), 18 days($E_2$) and 36 days($E_3$) separately, samples of the rostral superficial masseter muscle were obtained bilaterally and the proportion of type I, type IIA, type IIB, and type IIC fibers was determined and counted according to their histochemical activity of myosin ATPase (at pH 9.4, pH 4.6, and pH 4.2)and PAD staining. The obtained results were as follows : 1. The rostral superficial masseter muscle of the white rat contained approximately 47.5% type I fiber and 52.5% type II fiber. 2. Type I/ Type II ratio of molar-present side was increased significantly in the group E2 (18 days group) 3. Type IIA fiber was increased at molar-present side and decreased at molar-absent side in experimental groups.

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Ultrastructural and Cytochemical Studies on the Endometrial Surface Epithelial Cells of Guinea Pig During Estrous Cycle (發情週期에 따른 Guinea Pig의 子宮內膜 表層上皮細胞의 微細構造 및 細胞化學的 硏究)

  • Park, Choon K.;Kwan H. You;Young K. Deung;Lee, Choon K.;Ho S. Chung
    • The Korean Journal of Zoology
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    • v.25 no.1
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    • pp.9-28
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    • 1982
  • Cyclical changes in the fine structures of the surface epithelial, stroma and glandular cells of guinea pig endometrium during the estrous cycle were studied by transmission and scanning electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase, alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. The endometrial surface epithelium was pseudostratified columnar during estrus and meterstrus, and simple columnar during proestrus and diestrus. The characteristic features observed in these cells include increased nucleocytoplasmic ratio at proestrus, elongated shapes of both the nucleus and the entire cell, increased volume of the cytoplasm and cytoplasmic bulding into the lumen during estrus, and smaller surface epithelial cells during metestrus. 2. In the cytoplasm of surface epithelial cells, the numbers of mitochondria and free ribosomes were increased, and rough endoplasmic reticulum and Golgi complex appeared during estrus, and the degenerated cells, lipid droplets, multilamellated bodies and lysosomes appeared during diestrus. 3. During estrus, scanning electron microscopic observations of endometrial surface showed a regular arrangement with polygonal outlines of epithelial cells, distinct intercellular border, and bulged surface into the lumen, whereas flat surface and indistinct cell border were characteristic during meterstrus and diestrus. 4. Microvilli which aligned on the surface were longer and most abundant during estrus while short and aparse during other phases. 5. Cytochemical studies indicated that during metestrus acid phosphatase activities were localized in the microvilli and vacuoles, and alkaline phosphatase activities were significant around luminal surface and lateral cell membrane in the surface epithelial cells. ATPase activities were present on the microvilli and cell membrane during proestrus and estrus.

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