• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.9
• /
• pp.1274-1281
• /
• 2011

Dietary protein restriction affects lipid metabolism in rats. This study was performed to determine the effect of a low protein diet on hepatic lipid metabolism and insulin sensitivity in growing male rats. Growing rats were fed either a control 20% protein diet or an 8% low protein diet. Feeding a low protein diet for four weeks from 8 weeks of age induced a fatty liver. Expression of acetyl-CoA carboxylase, a key lipogenic enzyme, was increased in rats fed a low protein diet. Feeding a low protein diet decreased very low density lipoprotein (VLDL) secretion without statistical significance. Feeding a low protein diet down-regulated protein expression of microsomal triglyceride transfer protein, an important enzyme of VLDL secretion. Feeding a low protein diet increased serum adiponectin levels. We performed glucose tolerance test (GTT) and insulin tolerance test (ITT). Both GTT and ITT were increased in protein-restricted growing rats. Our results demonstrate that dietary protein restriction increases insulin sensitivity and that this could be due to low-protein diet-mediated metabolic adaptation. In addition, increased adiponectin levels may influences insulin sensitivity. In conclusion, dietary protein restriction induces a fatty liver. Both increased lipogenesis and decreased VLDL secretion has contributed to this metabolic changes. In addition, insulin resistance was not associated with fatty liver induced by protein restriction.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.8
• /
• pp.1316-1323
• /
• 2007

In order to induce high levels of protein secretion, we have constructed a recombinant plasmid, designated pBP244, into which was incorporated key components of the type-II See-dependent secretion system, including LepB (signal peptidase), SecA (ATPase), and SecB (chaperone). The biological activities of the LepB, SecA, and SecB components expressed from genes harbored by pBP244 appeared to play their normal roles. In order to evaluate the protein secretion, a pspA (Streptococcus $\underline{p}neumoniae\;\underline{s}urface\;\underline{p}rotein\;\underline{A}$) gene was cloned into pBP244, resulting in pBP438. S. typhimurium harboring pBP438 grown until the stationary phase, secreted a higher level of PspA into the culture supernatants than did the strain harboring pYA3494. The strain harboring pBP438 secreted a supernatant amount 1.71-fold, a periplasmic space amount 1.47-fold, and an outer membrane amount 1.49-fold higher than that of pYA3494. S. typhimurium ${\chi}8554$ kept the $Asd^+$ plasmid pBP244 and pBP438 for 60 generations in LB broth harboring DAP, thereby indicating that pBP244 and pBP438 were quite stable in the Salmonella strain.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.2
• /
• pp.144-151
• /
• 2014

Increasing the gene copy number has been commonly used to enhance the protein expression level in the yeast Pichia pastoris. However, this method has been shown to be effective up to a certain gene copy number, and a further increase of gene dosage can result in a decrease of expression level. Evidences indicate the gene dosage effect is product-dependent, which needs to be determined when expressing a new protein. Here, we describe a direct detection of the gene dosage effect on protein secretion through expressing the enhanced green fluorescent protein (EGFP) gene under the direction of the ${\alpha}$-factor preprosequence in a panel of yeast clones carrying increasing copies of the EGFP gene (from one to six copies). Directly examined under fluorescence microscopy, we found relatively lower levels of EGFP were secreted into the culture medium at one copy and two copies, substantial improvement of secretion appeared at three copies, plateau happened at four and five copies, and an apparent decrease of secretion happened at six copies. The secretion of EGFP being limiting at four and five copies was due to abundant intracellular accumulation of proteins, observed from the fluorescence image of yeast and confirmed by western blotting, which significantly activated the unfolded protein response indicated by the up-regulation of the BiP (the KAR2 gene product) and the protein disulfide isomerase. This study implies that tagging a reporter like GFP to a specific protein would facilitate a direct and rapid determination of the optimal gene copy number for high-yield expression.

• The Korean Journal of Physiology
• /
• v.20 no.2
• /
• pp.249-256
• /
• 1986

The present study was performed to investigate a possible influence of the adrenergic nervous system on pancreatic exocrine secretion stimulated by intraduodenal acid perfusion. Pancreatic secretion was collected in rats anesthetized with urethane after 24 hours fasting. The duodenal lumen was perfused (0.2 ml/min) with HCI solution in a concentration of 0.005, 0.01, 0.02, 0.05 or 0.1 N When the volume of panceratic juice secreted for IS min became constant phentolamine (1 mg/kg), $noradrenaline\;(10\;{\mu}g/kg),\;Propranolol\;(1\;mg/kg),\;and \;isoproterenol\;(1\;{\mu}g/kg)$ were administered through the jugular vein in bolus. The secretory volume and protein output were measured in the pancreatic juice collected for 15 min. 1) HCI, perfused intraduodenally in graded concentrations from 0.005 N to 0.1 N, increased the pancreatic secretory volume and protein output dose-dependently. 2) In the basal state as well as in the stimulated state by the duodenal acid perfusion, phentolamine increased the pancreatic secretory volume and protein output while propranolol inhibited the volume and protein output. 3) In the basal state, noradrenaline did not change the pancreatic secretory volume but increased the protein output while isoproterenol increased both of the secretory volume and the protein output. These results strongly suggest that ${\alpha}-adrenoceptors$ in the rat pancreas exert an inhibitory influence on the pancreatic exocrine secretion including volume and protein output in the basal state as well as in the stimulated state by the intraduodenal acid perfusion while ${\beta}-adrenoceptors$ play a stimulatory role in the pancreatic exocrine secretion. However, in the physiological situation, adrenergic excitation may stimulate the protein output through ${\beta}-adrenoceptors$ without change in the secretory volume in the rat pancreas.

• The Korean Journal of Zoology
• /
• v.3 no.1
• /
• pp.19-23
• /
• 1960

A method for determination of thyroid secretion rate in rabbit by means of radioactive iodine presented. After injection of radioactive iodine, in vivo determination so f radioactivity in thyroid gland were made during a 19 day-experimental period. In the same period blood samples were drawn and analyzed for protein-bound iodine (PBI) and for protein-bound radioactive iodine(PBI181). A rate constant for secretion of thyroid hormone was calculated from the disappearance rate of radioactive iodine in thyroid gland. The secretion rate of radioactive hormone iodine was calculated by multiplying this rate constant by the amount of radioactive iodine present in thyroid gland. Assuming that the specific radioactiveness of the circulating thyroid hormone and of the hormone just secreted were identical , thyroid secretion rate was calculated by the equation. {{{{ { Secreted hormone-iodine , gamma /hr} over { Secreted hormone-I^131, % dose/hr }= { PBI, ${\gamma}$/ml.Serum} over { PBI^131 , % dose/ml . Serum } }} The method presented consisted of measurements for series of independent criteria on thyroid function, and the resulting thyroid secretion rate was calculated by combination of those.

• Korean Journal of Microbiology
• /
• v.30 no.1
• /
• pp.60-64
• /
• 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.

• Korean Journal of Microbiology
• /
• v.30 no.2
• /
• pp.108-112
• /
• 1992

To investigate the expression and the secretion of heterologous proteins in yeast, we constructed an yeast secretion vector and produced a human secretory protein, .alpha.-1-antitrypsin (.alpha.-1-AT), from yeast cells. The secretion vector pGAT8 was constructed by inserting the signal sequence of yeast acid phosphatase gene (PH05) into the .alpha.1-AT expression vector pGAT6 which contained .alpha.-1-AT cDNA fused to GAL10-CYC1 promotor. The .alpha.-1-AT was produced efficiently in the yeast cells transformed with plasmid pGAT8, which was onfirmed both by the .alpha.-1-AT activity assay and by the immunoblot method using .alpha.-1-AT antibody. We also showed the secretion of .alpha.-1-AT into the culture media and into the periplasmic space by immunoblot.

• Molecules and Cells
• /
• v.26 no.3
• /
• pp.291-298
• /
• 2008

Human karyopherin ${\beta}3$, highly homologous to a yeast protein secretion enhancer (PSE1), has often been reported to be associated with a mediator of a nucleocytoplasmic transport pathway. Previously, we showed that karyopherin ${\beta}3$ complemented the PSE1 and KAP123 double mutant. Our research suggested that karyopherin beta has an evolutionary function similar to that of yeast PSE1 and/or KAP 123. In this study, we performed yeast two-hybrid screening to find a protein which would interact with karyopherin ${\beta}3$ and identified apolipoprotein A-I (apo A-I), a secretion protein with a primary function in cholesterol transport. By using in vitro binding assay, co-immunoprecipitation, and colocalization studies, we defined an interaction between karyopherin ${\beta}3$ and apo A-I. In addition, overexpression of karyopherin ${\beta}3$ significantly increased apo A-I secretion. These results suggest that karyopherin ${\beta}3$ plays a crucial role in apo A-I secretion. These findings may be relevant to the study of a novel function of karyopherin ${\beta}3$ and coronary artery diseases associated with apo A-I.

• Korean Journal of Veterinary Research
• /
• v.40 no.4
• /
• pp.677-681
• /
• 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.

• Proceedings of the Korean Biophysical Society Conference
• /
• 2001.06a
• /
• pp.56-56
• /
• 2001

Increasing evidence suggests that protein-protein interaction is essential in many biological processes including epithelial transport. In this report, we present the significance of protein interactions to HCO$_3$$^{-10}$ secretion in pancreatic duct cells. In pancreatic ducts HCO$_3$$^{-10}$ secretion is mediated by CFTR-activated luminal CUHCO$_3$$^{-10}$ exchange activity and HCO$_3$$^{-10}$ absorption is achieved by Na$^{＋}$-dependent mechanism including NHE3.(omitted)