• Title/Summary/Keyword: secretion

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Development of Protein Secretion System using Type III Secretion System of Salmonella (Salmonella Type III Secretion System을 이용한 단백질 분비시스템 개발)

  • Dinh, Le Tam Vo;Hong, Soon-Ho
    • KSBB Journal
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    • v.24 no.4
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    • pp.393-396
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    • 2009
  • New protein secretion system was developed using Type III Secretion System of Salmonella. N-terminal region of SlrP and SptP effector proteins were fused with TliA and EstA-P lipases by overlapping PCR. Lipase activity of Salmonella with SptP-TliA fusion system increased by 2.6 fold compare with wild type Salmonella strain. This result showed that lipase secretion via the T3SS would be a useful protein secretion machinery.

Roles of Gonadal Steroids on Exocrine Secretion of Isolated Perfused Rat Pancreas

  • Park, Hyung-Seo;Kim, Se-Hoon;Park, Hyoung-Jin;Lee, Mee-Young;Han, Young-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.4
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    • pp.217-221
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    • 2003
  • To clarify the roles of gonadal steroids on pancreatic exocrine secretion, effects of progesterone and estradiol-$17{\beta}$ on spontaneous and secretagogue-induced exocrine response of isolated perfused rat pancreas were investigated. Intra-arterial infusion of progesterone resulted in significant increase of the spontaneous pancreatic fluid and amylase secretion dose-dependently. However, estradiol-$17{\beta}$ did not exert any influence on spontaneous pancreatic exocrine secretion. Exogenous secretin, cholecystokinin (CCK), and acetylcholine markedly stimulated pancreatic fluid and amylase secretion. Progesterone initially enhanced secretin-induced amylase secretion, but this stimulatory response declined thereafter to basal value. Moreover, secretin-induced fluid secretion was not affected by infusion of progesterone. Therefore, initial increase of secretion-induced amylase secretion by progesterone seems to be a non-specific action by washout effect of secretin. Estradiol-$17{\beta}$ failed to change the secretin-induced fluid and amylase secretion. Both progesterone and estradiol-$17{\beta}$ did not exert any influence on CCK-induced fluid and amylase secretion. Acetylcholine-induced exocrine secretion of isolated perfused pancreas also was not affected by intra-arterial infusion of progesterone or estradiol-$17{\beta}$. It is concluded from the above results that progesterone could enhance the spontaneous pancreatic fluid and amylase secretion of isolated perfused rat pancreas through non-genomic shortterm action, and that these effects could be masked by more potent stimulants such as secretin, CCK, and acetylcholine.

Screening and Characterization of Secretion Signals from Lactococcus lactis ssp. cremoris LM0230

  • Jeong, Do-Won;Choi, Youn-Chul;Lee, Jung-Min;Seo, Jung-Min;Kim, Jeong-Hwan;Lee, Jong-Hoon;Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1052-1056
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    • 2004
  • A secretion signal sequence-selection vector (pGS40) was constructed based on an $\alpha$-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore $\alpha$-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgamo (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.

The regulatory mechanism of insulin like growth factor secretion by high glucose in mesangial cell: involvement of cAMP (Mesangial 세포에서 고포도당에 의한 insulin-like growth factor의 분비조절기전에 관한 연구: cAMP와의 관련성)

  • Heo, Jung-sun;Kang, Chang-won;Han, Ho-jae;Park, Soo-hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.563-571
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    • 2003
  • Dysfunction of mesangial cells has been contributed to the onset of diabetic nephropathy. Insulin like growth factors (IGFs) are also implicated in the pathogenesis of diabetic nephropathy. However, it is not yet known about the effect of high glucose on IGF-I and IGF-II secretion in the mesangial cells. Furthermore, the relationship between cAMP and high glucose on the secretion of IGFs was not elucidated. Thus, we examined the mechanisms by which high glucose regulates secretion of IGFs in mesangial cells. Glucose increased IGF-I secretion in a time- (>8 hr) and dose- (>15 mM) dependent manner (p<0.05). Stimulatory effect of high glucose on IGF-I secretion is predominantly observed in 25 mM glucose (high glucose), while 25 mM glucose did not affect cell viability and lactate dehydrogenase release. High glucose also increased IGF-II secretion. The increase of IGF-I and IGF-II secretion is not mediated by osmotic effect, since mannitol and L-glucose did not affect IGF-I and IGF-II secretion. 8-Br-cAMP mimicked high glucose-induced secretion of IGF-I and IGF-II. High glucose-induced stimulation of IGF-I and IGF-II secretion was blocked not by pertussis toxin but by SQ 22536 (adenylate cyclase inhibitor). Rp-cAMP (cAMP antagonist), and myristoylated protein kinase A (PKA) inhibitor amide 14-22 (protein kinase A inhibitor). These results suggest that cAMP/PKA pathways independent of Gi protein may mediate high glucose-induced increase of IGF-I and IGF-II secretion in mesangial cells. Indeed, glucose (>15 mM glucose) increased cAMP formation. In conclusion, high glucose stimulates IGF-I and IGF-II secretion via cAMP/PKA pathway in mesangial cells.

Putrescine and Cadaverine Enhance Insulin Secretion of Mouse Pancreatic ${\beta}$-cell Line

  • Park, Hyo-Eun;Kim, Jae-Young
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.193-200
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    • 2012
  • We examined the effects of polyamines such as putrescine and cadaverine on the biosynthesis and secretion of insulin in the mouse pancreatic ${\beta}$-cell line, MIN-6. Basal insulin secretion (BIS) and glucose-stimulated insulin secretion (GSIS) from the MIN-6 cells were significantly increased by 20 min- or 24 h-treatment with micromolar concentrations of polyamines. To determine whether the enhancement was due to increase of insulin production by polyamines, we investigated the insulin mRNA and protein production. Both insulin mRNA and protein production were found to be not significantly affected by the polyamine treatment. Next, we examined the expression of several transcription factors (TFs) related to insulin synthesis and secretion in order to identify upstream events responsible for the promotion of insulin secretion of MIN6 cells by polyamines. Of the 6 TFs tested, MafA was induced by treatment of polyamines. MafA mRNA and protein expressions increased with treatment of polyamines. Overall results suggest that cadaverine and putrescine promote the insulin secretion process rather than the insulin biosynthesis from MIN6 cells. Also MafA may be involved in the enhanced insulin secretion process. Further studies are needed to elucidate the underlying mechanisms for promotion of insulin secretion by polyamines.

ANP Inhibits Surfactant Secretion from Isoproterenol Stimulated Alveolar Type II Cells

  • Lee, Young-Man
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.65-70
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    • 1997
  • In order to investigate the effect of ANP on surfactant secretion from alveolar type II cell(AT II cell) during circulatory derangement in adult respiratory distress syndrome (ARDS), the secretion of surfactant from AT II cells was evaluated in purely isolated AT II cultures from rat lungs. For the simulation of sympathetic stimulation during circulatory derangement, primary AT II cultures were incubatedwith isoproterenol and IBMX. In this isoproterenol stimulated AT II cells, ANP were added in the media for the investigation of effect of ANP on surfactant secretion from AT II cells. For the evaluation of surfactant secretion, $[^3H]-methylcholine$ was incorporated and the level of radiolabelled choline chloride secreted from the cells was determined. As previously reported, isoproterenol and IBMX stimulated surfactant secretion from AT II cells. Isoproterenol showed synergistic increase of surfactant secretion with IBMX in AT II cells. In isoproterenol stimulated AT II cells, physiological level of ANP inhibited the secretion of surfactant in primary cultures of AT II cells. On the basis of these experimental it is suggested that, in association with ciculatory change during ARDS, increased secretion of ANP by the pulmonary edema, hypoxia and congestive heart heart failure might aggravate the symptoms of ARDS by reduction of surfactant secretion from AT II cells.

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Annexin I Stimulates Insulin Secretion through Regulation of Cytoskeleton and PKC Activity

  • Kang, Na-Na;Won, Jong-Hak;Park, Young-Min
    • Animal cells and systems
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    • v.13 no.1
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    • pp.17-23
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    • 2009
  • In previous studies, we found that Annexin I (Anx I) was co-secreted with insulin in response to glucose, and that extracellular Anx I stimulated the release of insulin via the Anx I binding site in rat pancreatic islets and the &-cell line. However, the role that Anx I plays in the insulin secretion was not established. Therefore, in this study, we evaluated the insulin secretion pattern in response to Anx I and the involvement of the cytoskeleton or PKC in Anx Istimulated insulin secretion in MIN6N8a cells. The peak time of insulin secretion in response to Anx I treatment corresponded with the second phase insulin secretion by glucose in the perifused pseudoislets. In addition, Anx I-stimulated insulin secretion was not affect by readily releasable pool depletion. Taken together, these findings indicate that Anx I treatment was associated with movement of the reserve pool of insulin. Furthermore, Anx I-stimulated insulin secretion was attenuated by treatment with a microfilament inhibitor, cytochalasin B, as well as by PKC down regulation. These results indicate that Anx I may be a regulator of second phase insulin secretion.

[ Sr2+ ] Stimulation of α-amylase and RNAse in Wheat Aleurone Layer

  • Kim, Tae-Wan
    • Korean Journal of Environmental Agriculture
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    • v.22 no.4
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    • pp.290-293
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    • 2003
  • To measure an effects of strontium on secretion of ${\alpha}$-amylase and RNase, wheat aleurone layers were isolated after the pre-incubation in a solution with or without 10 mM $SrCl_2$ or $CaCl_2$ for 3 days at $25^{\circ}C$ in the dark under aseptic conditions. The secretion of ${\alpha}$-amylase reached a maximum at 72 h after incubation. $Sr^{2+}$ induced more effectively secretion of ${\alpha}$-amylase than $Ca^{2+}$. The ${\alpha}$-amylase secretions by $Sr^{2+}$ or $Ca^{2+}$ ware about $2 (Ca^{2+})$ to $2.5 (Sr^{2+})$ fold higher than it without divalent ions, When aleurone layers were incubated without divalent ions, however, the ${\alpha}$-amylase was remarkably retained in the tissues. Total ${\alpha}$-amylase synthesis (ie. tissues + media) was slightly lowered by 10mM $SrCl_2$ addition. It seemed that the RNase secretion begins at 18 h after incubation. This meaned that the RNase secretion may process slower than ${\alpha}$-amylasee secretion. $Ca^{2+}$ effect on RNase secretion is stronger than $Sr^{2+}$ unlikely to ${\alpha}$-amylase. The secretion process is likely to be suddenly induced between 72 hand 96 h. These results suggested that the secretion was enhanced after the accumulation in aleurone layers.

Secretion of the cloned serratia marcescens nuclease in escherichia coli (Serratia marcescens nuclease의 escherichia coli에서의 분비)

  • 신용철;이상열;김기석
    • Korean Journal of Microbiology
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    • v.28 no.4
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    • pp.297-303
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    • 1990
  • Secretion of Serratia marcescens nuclease by E. coli harboring pNUC4 was investigated. 29.2, 54.2 and 16.6% of total nuclease were observed in culture medium, periplasm, and cytoplasm of E. coli, respectively. To investigate the secretion mechanism of Serratia nuclease by E. coli, secretion kinetics of nuclease was examined in the presences of sodium azide, and energy metabolism inhibitor; procaine, an exoprotein processing inhibitor; and chloramphenicol, a protein synthesis inhibitor. In the presence of sodium azide, periplasmic unclease was gradually decreased and the extracellular nyclease was linearly increased according to the incubation time. Similar results were obtained in presences of procaine and chloramphenicol. From these results, we concluded that two transport processes are involved in nuclease secretion: secretion of nuclease through the inner membrane is occurred by an energy-dependent process and probably requiring precusor processing: secretion of nuclease through outer membrane does not require energy, de novo protein synthesis, and precursor processing.

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Changes of spontaneous pancreatic exocrine secretion during the estrous cycle in rats (흰쥐에서 발정주기에 따른 자발적인 췌장외분비의 변화)

  • Park, Hyung-seo;Lee, Tae-im;Kim, Se-hoon;Park, Hyoung-jin;yang, Il-suk
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.677-681
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    • 2000
  • Since the role of female sexual hormones on pancreatic exocrine secretion was not fully understood, this study was investigated to clarify the difference of spontaneous pancreatic exocrine responses during the estrous cycle and the roles of ovarian hormones on pancreatic exocrine secretion in the anesthetized female rats. Pancreatic juice was collected from the sequential 15-min samples, and then fluid and protein secretion were measured from the collected samples. The stages of estrous cycle were defined by staining the vaginal smear. The spontaneous pancreatic fluid and protein secretion were significantly increased during the diestrus stage compare to the corresponding value during the estrus stage. In the ovariectomized rat, spontaneous pancreatic exocrine secretion was significantly decreased compare to the value of female rat during the diestrus stage and was restored by subcutaneous injection of progesterone (50 mg/kg). This results suggest that the spontaneous pancreatic exocrine secretion of female rat is fluctuated according to the estrous cycle and progesterone released from ovary could stimulate the spontaneous pancreatic exocrine secretion of female rat.

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