• Title/Summary/Keyword: intracellular transport

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Molecular chaperone as a sophisticated intracellular membership (세포내인자로서의 정교한 기능을 하는 molecular chaperone)

  • 권오유;송민호
    • Journal of Life Science
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    • v.8 no.2
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    • pp.226-226
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    • 1998
  • Discovery of molecular chaperone has stimulate cell biologists and thus made it possible to re-examine the processes whereby proteins achieve and maintain their functional conformations within living cells. the term ‘Molecular chaperone’ was first coined to describe one particular protein involved in the assembly of nucleosomes, but the term has now been extended to describe the function of a wide variety of proteins that assist protein transport across membranes, folding of nascent polypeptide, the assembly and disassembly of oligomeric structures, and the recovery or removal of proteins damaged by various environmental stresses including heat shock. Progress of molecular chaperone research is still limited by the lack of 3-dimensional structural information and detailed interacts with taget proteins in the cell. However, several laboratories around the world are attempting to extend our knowledge on the functions of molecular chaperone, and such efforts seem justified to finally provide the answers to the most burning questions shortly.

EFFECT OF GINSENG SAPONIN ON THE VASCULAR SMOOTH MUSCLE

  • Lee Kwang Soo
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.71-76
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    • 1980
  • Aortic strips were prepared from rabbits, and the tensions were maintained by administration of norepinephrine into the incubation chamber. The application of diol or triol induced relaxation of the aortic strip, as indicated by the decreased aortic tension. Triol, in a concentration of $30\;mg\%\;causes\;approximately\;50\%$ of muscle relaxation, whereas a similar degree of relaxation is induced by $50\;mg\%$ of diol. This indicates that both triol and diol cause relaxation of the aorta, but that triol is about $170\%$ more potent than diol. It is well established that blood-vessel smooth-muscle tone is regulated by the available intracellular $Ca^{++}$ concentration, which in turn is profoundly influenced by interaction of the cellular membrane and sarcoplasmic reticulum in the smooth muscle. Thus, any agent which modifies the smooth-muscle tone is expected to interfere with the $Ca^{++}$ binding or uptake of sarcolemma and sarcoplasmic reticulum. In the following experiments sarcoplasmic reticulum and sarcolemma were prepared from the ventricle of rabbit heart, and the active $Ca^{++}$ uptake by these cellular components was measured employing $Ca^{45}$ in the presence of triol and diol. It was found that the active $Ca^{++}$ uptake in the presence of ATP by sarcoplasmic reticulum was inhibited by both triol and diol. Panaxatriol, in a concentration of $80\;mg\;\%,$ inhibited $Ca^{++}$ uptake by $30\%,$ whereas panaxatriol in the same concentration inhibited uptake by $20\%.$ It is clear that triol is a more potent inhibitor of active $Ca^{++}$ transport in sarcoplasmic reticulum than diol. The $Ca^{++}$ binding of the cellular membrane was also studied employing Ca45 and milipore techniques. It was found that triol in a concentration of $80\;mg\;\%,$ decreased $Ca^{++}$ binding by $29\%.$ Diol in the same concentration decreased the binding by $17\%.$ It is clear that both triol and diol inhibit $Ca^{++}$ binding to the cellular membrane, but triol is approximately $180\%$ more potent than diol.

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Fumonisin $B_1$-induced Alteration of Sphingolipid Metabolism in $LLC-PK_1$ Cells ($LLC-PK_1$ 세포에서의 퓨모너신 $B_1$에 의해 유도된 스핑고리피드 대사)

  • Yoo, Hwan-Soo;Yun, Yeo-Pyo
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.789-796
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    • 1997
  • The purpose of this study was to determine the effect of sulfinpyrazone on fumonisin $B_1$-induced elevation of free sphingoid bases in LLC-$PK_1$ cells. Fumonis ins are a family of mycotoxins produced by the fungi Fusarium moniliforme which is common contaminant in corn. Fumonisins are also potent inhibiors of sphingosine and sphinganine N-acyltransferases (ceramide synthases), key enzymes in sphingolipid metabolism resulting in the elevation of free sphinganine. The cytosolic concentration of fumonisin B1 was known to be closely proportional to the elevation of free sphinganine in LLC-PK1 cells [Yoo, H.-S., Norred, W.P., Wang, E., Merrill, A.H., Jr., and Riley, R.T. (1992) Toxicol. Appl.Pharmacol. 114. 9-15]. Sulfinpyrazone, an anion transport inhibitor, reduced the elevated level of free sphinganine resulting from fumonisin B1 inhibition of de novo sphingolipid biosynthesis. Fumonisin B1 at a concentration of 20${\mu}$M showed approximately 120pmol/$10^6$ cells relative to 3-10pmol/$10^6$ cells in control cultures, and sulfinpyrazone at a concentration of 200${\mu}$M partially reversed the increased level of free sphinganine induced by fumonisin $B_1$ down to normal level after exposure to fumonisin $B_1$ for 8 to 24hr. However, the reduced effect of sulfinpyrazone on the fumonisin $B_1$-induced elevation of intracellular sphinganine was not shown after 24hr. Fumonisin $B_1$ exposure to LLC-PK1 cells for 36 and 48hr showed approximately 74 and 80pmol per $10^6$ cells relative to 82 and 76pmol,respectively, in fumonisin $B_1$ plus sulfinpyrazone-treated cultures. Sulfinpyrazone-induced less elevation of free sphinganine in confluent cells after exposure to fumonisin $B_1$ suggested that either sulfinpyrazone may block the availability of fumonisin $B_1$ to cells or act on the fumonisin $B_1$ interaction with ceramide synthase.

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Consensus channelome of dinoflagellates revealed by transcriptomic analysis sheds light on their physiology

  • Pozdnyakov, Ilya;Matantseva, Olga;Skarlato, Sergei
    • ALGAE
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    • v.36 no.4
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    • pp.315-326
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    • 2021
  • Ion channels are membrane protein complexes mediating passive ion flux across the cell membranes. Every organism has a certain set of ion channels that define its physiology. Dinoflagellates are ecologically important microorganisms characterized by effective physiological adaptability, which backs up their massive proliferations that often result in harmful blooms (red tides). In this study, we used a bioinformatics approach to identify homologs of known ion channels that belong to 36 ion channel families. We demonstrated that the versatility of the dinoflagellate physiology is underpinned by a high diversity of ion channels including homologs of animal and plant proteins, as well as channels unique to protists. The analysis of 27 transcriptomes allowed reconstructing a consensus ion channel repertoire (channelome) of dinoflagellates including the members of 31 ion channel families: inwardly-rectifying potassium channels, two-pore domain potassium channels, voltage-gated potassium channels (Kv), tandem Kv, cyclic nucleotide-binding domain-containing channels (CNBD), tandem CNBD, eukaryotic ionotropic glutamate receptors, large-conductance calcium-activated potassium channels, intermediate/small-conductance calcium-activated potassium channels, eukaryotic single-domain voltage-gated cation channels, transient receptor potential channels, two-pore domain calcium channels, four-domain voltage-gated cation channels, cation and anion Cys-loop receptors, small-conductivity mechanosensitive channels, large-conductivity mechanosensitive channels, voltage-gated proton channels, inositole-1,4,5-trisphosphate receptors, slow anion channels, aluminum-activated malate transporters and quick anion channels, mitochondrial calcium uniporters, voltage-dependent anion channels, vesicular chloride channels, ionotropic purinergic receptors, animal volage-insensitive cation channels, channelrhodopsins, bestrophins, voltage-gated chloride channels H+/Cl- exchangers, plant calcium-permeable mechanosensitive channels, and trimeric intracellular cation channels. Overall, dinoflagellates represent cells able to respond to physical and chemical stimuli utilizing a wide range of G-protein coupled receptors- and Ca2+-dependent signaling pathways. The applied approach not only shed light on the ion channel set in dinoflagellates, but also provided the information on possible molecular mechanisms underlying vital cellular processes dependent on the ion transport.

Polarization of THP-1-Derived Macrophage by Magnesium and MAGT1 Inhibition in Wound Healing

  • Mun Ho Oh;JaeHyuk Jang;Jong Hun Lee
    • Archives of Plastic Surgery
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    • v.50 no.4
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    • pp.432-442
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    • 2023
  • Background Macrophages play a major role in wound healing and prevent infection from the outside. Polarization conversion of macrophages regulates aspects of inflammation, and two macrophages, M1 (classically activated) and M2 (alternatively activated), exist at both ends of broad-spectrum macrophage polarization. Thus, we aimed to investigate whether macrophage polarization can be artificially regulated. To this end, MgSO4 and small-interfering RNA (siRNA) targeting magnesium transport 1 (MAGT1) were used to investigate the effects of intracellular magnesium (Mg2+) concentrations on the differentiation of macrophages in vitro. Methods THP-1 derived macrophages maintained in a culture medium containing 5 mM MgSO4 and siRNA to inhibit the expression of MAGT1. As comparative groups, THP-1 derived macrophages polarized into M1 and M2 macrophages by treatment with M1, M2 inducer cytokine. The polarization status of each group of cells was confirmed by cell surface antigen expression and cytokine secretion. Results We found that MgSO4 treatment increased CD163 and CD206, similar to the effect noted in the M2 group. The expression of CD80 and HLA-DR was increased in the group treated with MAGT1 siRNA, similar to the effect noted in the M1 group. Functional assays demonstrated that the group treated with MgSO4 secreted higher levels of IL-10, whereas the MAGT1 siRNA-treated group secreted higher levels of IL-6 cytokines. Additionally, the conditional medium of the Mg2+ treated group showed enhanced migration of keratinocytes and fibroblasts. Conclusion Mg2+ can help to end the delay in wound healing caused by persistent inflammation in the early stages.

Correlation between Rab3A Expression and Sperm Kinematic Characteristics

  • Seung-Ik Jang;Jae-Hwan Jo;Uwamahoro Claudine;Eun-Ju Jung;Woo-Jin Lee;Ju-Mi Hwang;Jeong-Won Bae;Dae-Hyun Kim;Jun Koo Yi;Jae Jung Ha;Dong Yep Oh;Woo-Sung Kwon
    • Development and Reproduction
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    • v.28 no.1
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    • pp.13-19
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    • 2024
  • Ras-related (Rab) proteins, integral members of the monomeric G-protein family, play a pivotal role in regulating intracellular vesicular transport. These proteins contribute to male reproductive processes, specifically in acrosome formation, exocytosis, and sperm motility. Although a prior study indicated a correlation between Rab3A and sperm motility, including motion kinematic parameters such as mean dance, this association has only been explored within a limited sample size. Therefore, further verification is required to confirm the correlation between Rab3A and sperm motility parameters. In the present study, Rab3A expression, sperm motility, and motion kinematic parameters were analyzed in 150 boar spermatozoa. Additionally, correlations between Rab3A expression and sperm kinematic characteristics were evaluated statistically. The results revealed significant associations between Rab3A protein expression levels and various motion kinematic parameters. Specifically, Rab3A levels exhibited positive correlations with average path velocity (p<0.05), mean amplitude of lateral head displacement (p<0.05), and curvilinear velocity (p<0.01). Consequently, it is proposed that Rab3A protein plays a crucial role in male fertility through its correlation with sperm kinematic characteristics, making it a potential marker for sperm motility-related assessments.

Effects of Cholesterol Diet and Exercise on Plasma and Liver Lipids, Platelet Aggregation and Erythrocyte Na Efflux in Rats (콜레스테롤 식이와 운동이 흰쥐의 체내 지질수준, 혈소판 응집 및 적혈구막 Na 유출에 미치는 영향)

  • Kang, Min-Sook;Kim, Young-Pyo;Kang, Young-Hee;Kang, Jung-Sook
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.381-390
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    • 2008
  • Present study was conducted to investigate the effects of exercise and cholesterol diet on plasma and liver lipids, platelet aggregation, erythrocyte Na efflux and liver index such as GOT and GPT using Sprague Dawley rats. Forty rats were divided into four groups and fed control or 0.5% cholesterol diet with and without treadmill for six weeks. The final body weight of group fed cholesterol diet with exercise was somewhat decreased compared with group fed cholesterol diet without exercise. L.W/B.W ratio was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise tended to decrease this ratio. Plasma total cholesterol was significantly increased and HDL-cholesterol was decreased in groups fed cholesterol diet (p < 0.01). Plasma triglyceride was significantly decreased in groups fed cholesterol diet compared with groups fed control diet (p < 0.01). Plasma triglyceride of groups with exercise was significantly decreased compared with their non exercising counterparts regardless diet (p < 0.05). Liver total cholesterol and triglyceride was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise did not affect on these levels. Na-K ATPase was somewhat decreased in groups fed cholesterol diet, and exercise tended to recover the reduced Na-K ATPase. Na passive transport was significantly decreased in group fed control diet without exercise and significantly increased in group fed cholesterol diet with exercise, there were significant differences between groups (p < 0.05). There were no differences in total Na efflux and intracellular Na among groups, and total Na efflux was not correlated with intracellular Na. Hematocrit was significantly lower (p < 0.05) in group fed cholesterol diet without exercise compared with other groups. Platelet aggregation in the initial slope and the maximum was increased in groups fed cholesterol diet, but not statistically significant. Exercise especially increased the initial slope of aggregation. Plasma GOT and GPT was significantly increased in groups fed cholesterol diet (p < 0.01), and exercise in group fed cholesterol diet significantly decreased both GOT and GPT compared with the non exercising counterpart (p < 0.01). This study showed that cholesterol diet increases plasma and liver lipids and GOT and GPT, and exercise improves plasma and liver lipid profile and liver index of GOT and GPT preventing fatty liver.

Rab Effector EHBP1L1 Associates with the Tetratricopeptide Repeat Domain of Kinesin Light Chain 1 (Kinesin Light Chain 1 (KLC1)의 Tetratricopeptide Repeat (TPR) 도메인과 Rab effector, EHBP1L1의 결합)

  • Jeong, Young Joo;Park, Sung Woo;Kim, Sang-Jin;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.30 no.1
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    • pp.10-17
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    • 2020
  • Kinesin-1 is microtubule-dependent plus-end direct molecular motor protein essential for intracellular transport. It is a member of the kinesin superfamily proteins (KIFs) which transport cargo, including organelles, vesicles, neurotransmitter receptors, cell-signaling molecules, and protein complexes through interaction between its light chain subunit and the cargo. Kinesin light chain 1 (KLC1) is a non-motor subunit that associates with the kinesin heavy chain (KHC). Although KLC1 interacts with many different adaptor proteins and scaffolding proteins, its binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1, and found an interaction between KLC1 and EH domain-binding protein 1 like 1 (EHBP1L1). EHBP1L1 bound to the region containing all six TPR repeats of KLC1 and did not interact with KIF5B (a motor protein of kinesin 1) or KIF3A (a motor protein of kinesin 2) in the yeast two-hybrid assay. The carboxyl-terminus of the coiled-coil domain of EHBP1L1 is essential for interaction with KLC1. However, another EHBP1L1 isoform, EHBP1, did not interact with KLC1 in the yeast two-hybrid assay. KLC1 interacted with GST-EHBP1L1 and its coiled-coil domain but not with GST only. When co-expressed in HEK-293T cells, EHBP1L1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B but not KIF3A. These results suggest that kinesin 1 motor protein may transport EHBP1L1-associated cargo in cells.

Some Observations on the Fine Structure of Vacuolar Apparatus Affected by Dehydrocholic acid, Cholesterol and Phosphatidylcholine (담즙산과 cholesterol 및 phosphatidylcholine이 vacuolar apparatus에 미치는 영향에 관한 미세구조적 관찰)

  • Shin, Young-Chul
    • Applied Microscopy
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    • v.26 no.4
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    • pp.465-477
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    • 1996
  • The influence of dehydrocholic acid, cholesterol and phosphstidylcholie to the fine structure of vacuolar apparatus was investigated to better understand the mechanism of intracellular transport of bile constituents in the hepatocytes of rats. The cis Golgi cisterns faced toward the bile canaliculi both in normal and experimental groups. In the hepatocytes from the rats of experimental groups, the primary organic solutes in bile influence the Gogi apparatus, ER and lysosome in the way of increase, cisternal dilation or budding to form the vacuoles. In the dehydrocholic acid group, the cis Golgi cisterns appeared to be sacculated and showed buds, which were probably separated to be vacuoles. Some of the vacuoles appeared to be fused to the bile canaliculi. In the cholesterol and phosphatidylcholine groups, the Golgi cisterns appeared to be dilated and lysosomes were increased in the vicinity of bile canaliculi. The cis Golgi cisterns showing linear saccular fashions were occasonally observed. The increase of lysosomes were more predominant in the cholesterol group. The evidence suggests that dehydrocholic acid is mainly transported through the ER and cis Golgi cisterns, and cholesterol and phosphatidylcholine are mainly transported through the ER and lysosomes via the trans Golgi cisterns, but the cholesterols are frequently transported via the lysosomes.

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Regulation of S100G Expression in the Uterine Endometrium during Early Pregnancy in Pigs

  • Choi, Yo-Han;Seo, Hee-Won;Shim, Jang-Soo;Kim, Min-Goo;Ka, Hak-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.44-51
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    • 2012
  • Calcium ions play an important role in the establishment and maintenance of pregnancy, but molecular and cellular regulatory mechanisms of calcium ion action in the uterine endometrium are not fully understood in pigs. Previously, we have shown that calcium regulatory molecules, transient receptor potential vanilloid type 5 (TRPV6) and calbindin-D9k (S100G), are expressed in the uterine endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner, and that estrogen of conceptus origin increases endometrial TRPV6 expression. However, regulation of S100G expression in the uterine endometrium and conceptus expression of S100G has been not determined during early pregnancy. Thus, we investigated regulation of S100G expression by estrogen and interleukin-$1{\beta}$ (IL1B) in the uterine endometrium and conceptus expression of S100G during early pregnancy in pigs. We obtained uterine endometrial tissues from day (D) 12 of the estrous cycle and treated with combinations of steroid hormones, estradiol-$17{\beta}$ ($E_2$) and progesterone ($P_4$), and increasing doses of IL1B. Real-time RT-PCR analysis showed that $E_2$ and IL1B increased S100G mRNA levels in the uterine endometrium, and conceptuses expressed S100G mRNA during early pregnancy, as determined by RT-PCR analysis. To determine if endometrial expression of S100G mRNA during the implantation period was affected by the somatic cell nuclear transfer (SCNT) procedure, we compared S100G mRNA levels in the uterine endometrium from gilts with SCNT-derived conceptuses with those from gilts with conceptuses derived from natural mating on D12 of pregnancy. Real-time RT-PCR analysis showed that levels of S100G mRNA in the uterine endometrium from gilts carrying SCNT-derived conceptuses was significantly lower than those from gilts carrying conceptuses derived from natural mating. These results showed that S100G expression in the uterine endometrium was regulated by estrogen and IL1B of conceptus origin, and affected by the SCNT procedure during early pregnancy. These suggest that conceptus signals regulate S100G, an intracellular calcium transport protein, for the establishment of pregnancy in pigs.