• 미생물학회지
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• 제28권4호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제10권2호
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• pp.233-239
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• 1997

A mammary epithelial cell line (MAC-T) established as a model for lactation was utilized to identify and characterize effects of various hormones upon insulin-like growth factor binding protein secretion. Ligand and immunoblot analyses of conditioned media indicated that insulin-like growth factor binding protein-2 was secreted by MAC-T cells. Insulin-like growth factor-I stimulated insulin-like growth factor binding protein-2 secretion in a dose-dependent manner, but prolactin and bovine somatotropin did not alter insulin-like growth factor binding protein-2 secretion. Insulin increased and cortisol decreased insulin-like growth factor binding protein-2 secretion. Effects of insulin-like growth factor-I on insulin-like growth factor binding protein-2 secretion support previous studies using primary cultures of bovine mammary cells and bovine fibroblasts. Effects of cortisol and insulin on insulin-like growth factor binding protein-2 secretion may be explained by changes in protein synthesis. In addition, supraphysiological doses of insulin can cross-react with the insulin-like growth factor-I receptor and stimulate insulin-like growth factor binding protein-2 secretion. MAC-T cells provide a model system to study mechanisms that regulate local insulin-like growth factor-I bioactivity.

• Journal of Nutrition and Health
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• 제27권9호
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• pp.910-923
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• 1994

The effects of individual fatty acids, differing in their degree of unsaturation(18:0, 18:1, 18:2 and 18:3) on the biosynthesis and secretion and lipids were investigated in Hep-G2 cells. Synthesis of apolipoprotein was measured by the incorporation of 3H-leucine into apolipoprotein(d<1.21g/ml) and synthesis of lipids was measured by the incorporation of 3H-glycerol and 14C-acetate into various lipid classes. Inclusion of 1.0mM of each fatty acids into the culture medium significantly increased the synthesis of total apolipoprotein and Apo B(p<0.05). However, addition of fatty acid did not affect the synthesis of cellular and medium protein. Among different fatty acids tested, oleic acid had the greatest effect on Apo B synthesis. While stearic, linoleic and linolenic acid, all had similar effects. The secretion of triglyceride into the medium markedly increased in all fatty acid groups being 5-6 times over the albumin control. The triglyceride secretion was the highest int he oleic acid group. The secretion of phospholipid and cholesterol also increased with triglyceride output. A positive relationship existed between the output of lipoprotein-triglyceride and Apo B. Since the synthesis of Apo B was significantly increased when various fatty acids were included into the culture medium, part of the apparently stimulated synthesis of the apolipoprotein may be in response to the increased formation and secretion of lipoprotein lipids.

• 한국동물학회지
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• 제30권2호
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• pp.193-209
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• 1987

Patterns of amylase secretion in mouse pancreatic fragments were studied over a period of time after the tissue was stimulated by acetyicholine and MNNG. MNNG is known to activate guanylate cyclase and thus increase the cGMP concentration in the pancreatic acinar cell. These amylase secretion patterns were studied to investigate the role of cGMP in reaction cascade during secretion response of the tissues stimulated by acetyicholine. Cellular response of amylase secretion in the pancreas by acetyicholine was divided into two phases. During the first phase, zymogen granules which had existed in the cells were secreted by the action of $Ca^2$+ and calmodulin immediately after secretagogue administration, this being known as the initial response. When the tissue was stimulated by acetylcholine in a $Ca^2$+-deficient medium or one containing trifluoperazine as a calmodulin antagonist, this initial response was reduced. In the second phase, newly formed zymogen granules were secreted as sustained response after protein synthesis was triggered by secretagogue. This response was provoked by an activation of protein kinase C. When either cycloheximide as a protein synthesis inhibitor or dibucaine as a protein kinase C inhibitor were added to the incubation medium, this sustained response was remarkablely depressed in the pancreatic fragments stimulated with acetylcholine. In the pancreatic acinar cell, phosphatidylinositol turnover plays an important role in the secretion response and hexachlorocyclohexane inhibits this phosphatidylinositol turnover. The pancreatic tissue treated with the hexachlorocyclohexane exhibited inhibition on both initial and sustained responses of amylase secretion by acetylcholine. MNNG also accelerated amylase secretion from the tissue gradually along incubation time. The 22 minutes fraction of the pancratic secretion after administration of both acetylcholine and MNNG showed higher amylase activity than the neighboring fractions. Guanylate cyclase potentiated the sustained response. Even if it is experimented with an indirect method, guanylate cyclase was found responsible for activation of the sustained response of a step prior to the action of protein kinase C. As conclusion, it was considered that amylase secretion in mouse pancreatic fragments stimulated by acetylcholine is a three phasic response.

• Toxicological Research
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• 제9권2호
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• pp.195-206
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• 1993

The expression of proenkephalin (proENK) mRNA and Met-enkephalin (ME) secretion in C6 rat glioma cells and bovine adrenal medullary chromaffin (BAMC) cells were elucidated in the present study. The levels of proENK mRNA and ME secreted into the media in BAMC cells were measured in the presence of cycloheximide and 12-tetrade-canoylphorbol-13-acetate (TPA). Cycloheximide (20 nM) abolished the induction of proENK mRNA expression, protein synthesis and ME secretion by TPA (1nM), indicating that de novo protein synthesis was necessary for proENK gene expression and ME secretion.

• 한국동물학회지
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• 제35권3호
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• pp.271-276
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• 1992

20-hydroxvecdysone (20-HE) seems to be related in the regulation of vitellogenesis in Drosophifa sp. (robwta species sroup). Although yolk proteins (YPs) synthesis does not occur at a high rate in fat body cells of one daw-old female after eclosion, application of 20-HE to isolated abdomens deprived of anterior endocrine glands stimulated the synthesis and secretion of YPs into the hemolvmph. An injection of 0.3 $\mul$ of a 10-s M 20-HE was sufficient topromote synthesis and secretion of YPs in isolated abdomens. The response of isolated automens to hormones was first detected between 2 hr and 3 hr after treaDent of 10-s M 20-HE. Transcript analysis showed that the effect of 20-HE on yolk protein synthesis was mediated at the level of transcription.

• 대한의생명과학회지
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• 제18권3호
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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.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.1977-1988
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• 2015

β-1,3-glucanosyltransferases play essential roles in cell wall biosynthesis in yeast. Kluyveromyces lactis has six putative β-1,3-glucanosyltransferase genes. KlGAS1-1 and KlGAS1-2 are homologs of Saccharomyces cerevisiae gene GAS1. RT-qPCR indicated the transcription level of KlGAS1-1 was significantly reduced while heterologous protein (thermostable xylanase B) secretion was enhanced during medium optimization. To evaluate if these two events were related, and to improve xylanase B secretion in K. lactis, we constructed KlGAS1-1 and KlGAS1-2 single deletion strains and double deletion strain, respectively. KlGAS1-1 gene deletion resulted in the highest xylanase B activity among the three mutants. Only the double deletion strain showed morphology similar to that of the GAS1 deletion mutant in S. cerevisiae. The two single deletion strains differed in terms of cell wall thickness and xylanase B secretion. Transcription levels of β-1,3-glucanosyltransferase genes and genes related to protein secretion and transport were assayed. The β-1,3-glucanosyltransferase genes displayed transcription complementation in the cell wall synthesis process. KlGAS1-1 and KlGAS1-2 affected transcription levels of secretion- and transport-related genes. Differences in protein secretion ratio among the three deletion strains were associated with changes of transcription levels of secretion- and transport-related genes. Our findings indicate that KlGAS1-1 deletion is an effective tool for enhancing industrial-scale heterologous protein secretion in K. lactis.

• 미생물학회지
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• 제30권1호
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• pp.60-64
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• 1992

The secretion vector with promoter and signal sequence region of neutral protease gene (npr) from Bacillus amyloliquefaciens was constructed by the technique of polymerase chain reaction (PCR). A unique restriction iste was introduced into the 3' of the signal coding region by the synthesis of PCR primer. To demonstrate the function of cloned promoter and signal sequence, we used the E. coli .betha.-lactamase structural gene as a foreign gene. The signal sequence of .betha.-lactamase gene was deleted by Bal31 exonuclease and only mature region was introduced into the secretion vector. Bacillus subtilis cells transformed by the recombinant vector synthesized the fusion protein and were also capable of removing the signal peptide from the original fusion protein, as judged by the assay of .betha.-lactamase activity and secretion into the growth medium by western blotting.

• The Korean Journal of Physiology and Pharmacology
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• 제4권2호
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• pp.129-135
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• 2000

Previously, we have reported that ${\rho}-chlorophenylalanine$ (PCPA), a serotonin depletor, profoundly increased pancreatic fluid and bicarbonate secretion but remarkably inhibited pancreatic amylase secretion in anesthetized rats. The present study was performed to verify the detailed effects of PCPA on pancreatic amylase synthesis that is directly related to amylase exocrine secretion. PCPA significantly decreased pancreatic RNA and protein contents as well as the amylase activity. However, pancreatic DNA content, trypsin and chymotrypsin activities were not influenced by the treatment of PCPA. The rate of pancreatic amylase synthesis, which was assessed by the amount of incorporated $[^{35}S]-methionine$ into amylase for 1 h, was also significantly decreased by 44% in PCPA-treated rats. In order to determine whether the PCPA-induced decrease of amylase synthesis resulted from change in the level of amylase mRNA, Northern blot analysis was performed. The mRNA expression level of amylase was also decreased by 48% in the PCPA-treated rats, indicating that the inhibitory effect of PCPA on the synthesis of pancreatic amylase was mainly regulated at a step prior to translation. It was also revealed in SDS-polyacrylamide gel electrophoresis that the qualitative change of amylase was induced by PCPA. The 54 KDa amylase band seems to be degraded into small molecular weight protein bands in PCPA-treated rats, suggesting that the PCPA- induced decrease of amylase may be partly attributed to the degradation of synthesized amylase.