Two alternative pathways for methionine biosynthesis are known in Corynebacterium glutamicum: one involving transsulfuration (mediated by metB and metC) and the other involving direct sulfhydrylation (mediated by metY). In this study, MetB (cystathionine ${\gamma}-synthase$) and MetY (O-acetylhomoserine sulfhydrylase) from C. glutamicum were purified to homogeneity and the biochemical parameters were compared to assess the functional and evolutionary importance of each pathway. The molecular masses of the native MetB and MetY proteins were measured to be approximately 170 and 280 kDa, respectively, showing that MetB was a homotetramer of 40-kDa subunits and MetY was a homohexamer of 45-kDa subunits. The $K_m$ values for the O-acetylhomoserine catalysis effected by MetB and MetY were 3.9 and 6.4 mM, and the maximum catalysis rates were $7.4\;(k_{cat}=21\;S^{-1})\;and\;6.0\;(k_{cat}=28\;S^{-1})\;{\mu}mol\;mg^{-1}\;min^{-1}$, respectively. This suggests that both MetB and MetY can be comparably active in vivo. Nevertheless, the $K_m$ value for sulfide ions by MetY was 8.6mM, which was too high, considering the physiological condition. Moreover, MetB was active at a broad range of temperatures $(30\;and\;65^{\circ}C)$ and pH (6.5 and 10.0), as compared with MetY, which was active in a range from 30 to $45^{\circ}C$ and at pH values from 7.0 to 8.5. In addition, MetY was inhibited by methionine, but MetB was not. These biochemical data may provide insight on the role of the parallel pathways of methionine biosynthesis in C. glutamicum with regard to cell physiology and evolution.
Journal of the Korean Society of Food Science and Nutrition
/
v.23
no.3
/
pp.429-435
/
1994
The effect of methionine (Met) supplementation on glutathione peroxidase(GSHPx) activity in young and 14 month-old rat and mice was investigated. GSHPx activity was more enhanced by methionine supplementation in young rats when selenium (Se) was given as selenite than given in the form of selenomethione (Se-Met). However, GSHPx activity was not influenced by Met supplementation in the old rats. When diets were low in Se, the biopotency of ht eenzyme by Met was facilitated. No significant differences in GSHPx activity was observed with Met supplement in growing mice when Met was given 0.3% and 0.8% iin the diet at high levels of Se (2 ppm). The peak GSHPx in liver and kidney occurred at day 18, thereafter it decreased. Particularly, the liver GSHPx at day 18 increased 4.2 times than that at day 4 by 0.5% Met supplementation, while the unsupplemented group remained only 2.5 times increase. It is considered that in some tissues Met requirement may be met by Se-Met when rats were fed a diet suboptimal in Met. In addition, at lower levels of Se the utilization of Se is more enhanced by Met than at higher levels of dietary Se. Therefore, GSHPx activity may be influenced greatly by Met status along with dietary Se.
Journal of the Korean Society of Food Science and Nutrition
/
v.26
no.1
/
pp.109-115
/
1997
This study was designed to investigate the effects of methionine(Met) on the activities of brain lipid peroxidation related enzymes in ethanol administrated rats of selenium(Se) deficiency. Male Sprague-Dawley rats were fed Se deficiency diets containing one of the three levels of Met (0, 3, 9g/kg diet) and ethanol(2.5g/kg of body weight) was administrated as 25v/v% ethanol treated groups orally. The rats sacrificed after 5 and 10 weeks of feeding periods. Alcohol dehydrogenase activity was increased in ethanol treated groups and was higher Met normal group than Met deficiency and excessive groups at 5 and 10 weeks dieting. Aldehyde dehydrogenase activity was decreased in ethanol treated groups and significantly decreased in Met deficiency group. Monoamine oxidase activity in brain was increased in ethanol treated groups and was predominently increased in Met deficiency groups. Superoxide dismutase and glutathione peroxidase activities were decreased in ethanol treated groups and tended to increase in proportion to level of dietary methionine. Glutathione S-transferase and catalase activities and lipid peroxide content were increased by ethanol administration and were higher Met deficiency group than normal and excessive groups.
Cho, Sung Ho;Kim, Jong In;Kim, Hyun Su;Park, Sung Dal;Jang, Kang Won
Journal of Chest Surgery
/
v.50
no.3
/
pp.153-162
/
2017
Background: The mesenchymal-epithelial transition factor (MET) receptor can be overexpressed in solid tumors, including small cell lung cancer (SCLC). However, the molecular mechanism regulating MET stability and turnover in SCLC remains undefined. One potential mechanism of MET regulation involves the C-terminus of Hsp70-interacting protein (CHIP), which targets heat shock protein 90-interacting proteins for ubiquitination and proteasomal degradation. In the present study, we investigated the functional effects of CHIP expression on MET regulation and the control of SCLC cell apoptosis and invasion. Methods: To evaluate the expression of CHIP and c-Met, which is a protein that in humans is encoded by the MET gene (the MET proto-oncogene), we examined the expression pattern of c-Met and CHIP in SCLC cell lines by western blotting. To investigate whether CHIP overexpression reduced cell proliferation and invasive activity in SCLC cell lines, we transfected cells with CHIP and performed a cell viability assay and cellular apoptosis assays. Results: We found an inverse relationship between the expression of CHIP and MET in SCLC cell lines (n=5). CHIP destabilized the endogenous MET receptor in SCLC cell lines, indicating an essential role for CHIP in the regulation of MET degradation. In addition, CHIP inhibited MET-dependent pathways, and invasion, cell growth, and apoptosis were reduced by CHIP overexpression in SCLC cell lines. Conclusion: C HIP is capable of regulating SCLC cell apoptosis and invasion by inhibiting MET-mediated cytoskeletal and cell survival pathways in NCI-H69 cells. CHIP suppresses MET-dependent signaling, and regulates MET-mediated SCLC motility.
A layer experiment was conducted to determine the effects of supplementary methionine chelates (Cu, Zn and Mn), individual or in combination, on laying performance, eggshell quality, gizzard erosion, and IgG level of serum for 8 weeks. Five hundred 96-wk-old force molted ISA Brown layers were assigned to five dietary treatments. Basal diet was formulated to meet or exceed the nutrients requirements listed in NRC (1994). Five experimental diets were control, Zn-methionine chelate (Zn-Met) supplemented, Cumethionine chelate (Cu-Met) supplemented, Zn-Mn-methionine chelate (Zn-Mn-Met) supplemented and Zn-Mn-Cu-Met supplemented diet. Each treated diet was supplemented with respective mineral(s) at the level of 100 ppm in the form of methionine chelate. Egg production was increased by Cu-Met supplementation but decreased by Zn-Met supplementation. Egg weight was significantly (p<0.05) lower in Cu-Met treatment than those of the control and Zn-Met treatment. Specific gravity of eggs and eggshell strength were highest and soft egg production was lowest in Cu-Met treatment. Gizzard erosion index was significantly increased by supplementation of Cu-Met, Zn-Mn-Met or Zn-Mn-Cu-Met. Zinc content in liver significantly increased by Zn-Met, but not by Zn-Mn-Cu-Met treatment. In conclusion, 100 ppm Cu in Cu-Met chelate improved laying performance and eggshell quality but also increased gizzard erosion index. Supplementation of Zn-Met or its combination with other mineral chelates had no beneficial effects on laying performance and eggshell quality.
In this study, we focused on myomoduline A (MMA) released from the central nervous system of Aplysia kurodai. The primary structure of MMA is Pro-Met-Ser-Met-Leu-Arg-Leu-$NH_2$. This peptide is the same as that of the myomodulin family peptide found in other mollusks. The purified MMA showed a modulating activity of phasic contraction on the anterior byssus retractor muscle (ABRM) of Mytilus edulis. In order to study the relationship between the structure and biological activity of MMA, we synthesized MMA, Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, Des[$Pro^1,Met^2,Ser^3$]-MMA, and MME. The amino acid sequences of Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, and Des[$Pro^1,Met^2,Ser^3$]-MMA were Met-Ser-Met-Leu-Arg-Leu-$NH_2$, Ser-Met-Leu-Arg-Leu-$NH_2$, and Met-Leu-Arg-Leu-$NH_2$, respectively. MMA and synthetic peptides were tested on ABRM in M. edulis as well as muscle preparations in Achatina fulica. At $1{\times}10^{-8}$ M or lower, MMA showed a potentiating effect on phasic contraction of the ABRM, but this peptide had an inhibitory effect at $1{\times}10^{-6}$ M or higher. Both MMA and its analogs stimulated a contractile response on the crop and a relaxed catch-relaxing response on the penial retractor muscle of A. fulica. These results suggest that Met-Leu-Arg-Leu-$NH_2$ in MMA is the minimum structure required for the regulation of the contraction of ABRM, as well as the reproductive and digestive activities of mollusks.
The multiple ear and tiller line (MET), which was selected from a local maize collection and has been selected for earliness by selfing for five generations, was compared its plant characteristics with four U.S. leading hybrids. 1. The general plant growth was a little retarded by poor soil fertility and structure. 2. The fresh weight of the MET line was about 97.8% of the U.S. check varieties. 3. The dry weight of the MET line was also about 94.5% of the check U.S. hybrids (1,695 kg / 10a.). 4. The tillering and eating habits of the MET line had contributed toward increasing fresh and dry weight of the MET line. 5. The average plant height of the MET line was much less than those of the check varieties, due to the selfing depression manifested by the MET line, (234.7cm. of the MET vs 250.7cm. of the check) 6. Apparent differences of disease resistance was observed among varieties studied. The degree of plants infected with the black stripe mosaic virus was over 20% for the check U.S. hybrids, while less than 5% of the MET line was infected with the disease. 7. The grain yield per 10 a of the MET line was far below the check hybrids due to the inbreeding depression of the MET line. However, the kernel number per unit area of the MET line with small size kernel was much greater than that of the check hybrids. The 100 kernel weight of the MET line was around 10 grams. 8. The average fresh and dry matter weight of the three ($sorghum{\times}sudan$ grass) varieties was very much alike with those of the MET line. 9. In conclusion, the use of the MET line for silage production may have dual adventages because of its low seed price and of its small size kernels, which may reduce the amount of seeds required for planting in an unit area.
Purpose: The aim of the present study was to elucidate the clinicopathological significance and diagnostic accuracy of immunohistochemistry (IHC) for determining the mesenchymal epidermal transition (c-MET) expression in patients with gastric cancer (GC). Materials and Methods: The present meta-analysis investigated the correlation between c-MET expression as determined by IHC and the clinicopathological parameters in 8,395 GC patients from 37 studies that satisfied the eligibility criteria. In addition, a concordance analysis was performed between c-MET expression as determined by IHC and c-MET amplification, and the diagnostic test accuracy was reviewed. Results: The estimated rate of c-MET overexpression was 0.403 (95% confidence interval [CI], 0.327~0.484) and it was significantly correlated with male patients, poor differentiation, lymph node metastasis, higher TNM stage, and human epidermal growth factor receptor 2 (HER2) positivity in IHC analysis. There was a significant correlation between c-MET expression and worse overall survival rate (hazard ratio, 1.588; 95% CI, 1.266~1.992). The concordance rates between c-MET expression and c-MET amplification were 0.967 (95% CI, 0.916~0.987) and 0.270 (95% CI, 0.173~0.395) for cases with non-overexpressed and overexpressed c-MET, respectively. In the diagnostic test accuracy review, the pooled sensitivity and specificity were 0.56 (95% CI, 0.50~0.63) and 0.79 (95% CI, 0.77~0.81), respectively. Conclusions: The c-MET overexpression as determined by IHC was significantly correlated with aggressive tumor behavior and positive IHC status for HER2 in patients with GC. In addition, the c-MET expression status could be useful in the screening of c-MET amplification in patients with GC.
The objective of the current study was to evaluate the effect of supplementation of either L-methionine (L-Met) or DL-methionine (DL-Met) to diets of starter pigs on nitrogen (N) balance, metabolism, and serum amino acid profile. Eighteen crossbred ($Duroc{\times}Landrace{\times}Yorkshire$) barrows weighing $15.45{\pm}0.88kg$ were randomly allotted to 1 of 3 diets with 6 pigs per treatment. The diets included a basal diet (Met-deficient diet) containing 0.24% standardized ileal digestibility Met with all other essential nutrients meeting the pig's requirements. The other two diets were produced by supplementing the basal diet with 0.12% DL-Met or L-Met. The experiment lasted for 18 days, consisting of a 13-day adaptation period to the diets followed by a 5-day experimental period. Pigs were fed ad libitum and free access to water throughout the experiment. Results showed that the supplementation of either L-Met or DL-Met improved N retention, and serum methionine concentration, and decreased N excretion compared with basal diet (p<0.01). The N retention of pigs fed diets supplemented with the same inclusion levels of DL-Met or L-Met were not different (p>0.05). In conclusion, on equimolar basis DL-Met and L-Met are equally bioavailable as Met sources for starter pigs.
Silage yield and nutritional values of a locally collected Korean corn line with high tillering and earing characteristics were investigated at plant densities from 2778 to 8333 plants/ l0a. This line and a non-tillering and nonprolific corn as a check were grown at four plant densities and at three planting dates. The Korean local line abbreviated as MET had the highest dry matter per l0a. The highest dry matter of the MET line compared with the check hybrid was due to the highest dry leaf weight of the MET line. The highest dry leaf weight of the MET line was due to the increased number of tillers of the MET line. Other plant parts such as ear weight, kernel weight and cob weight of the MET line were lower than those of the check hybrid. The dry husk weight per l0a of the MET line was higher than that of the check hybrid, probably due to the increased number of ears in the MET line. The total embryo production per 10 a of the MET line was significantly higher than that of the check hybrid. The increased portion of embryo of the MET line is probably responsible for the higher TON values of the MET line. No interaction between variety x planting dates or planting density was found, indicating that the MET line and check hybrid were both the same in effects of planting dates and densities. Both line and hybrid showed the highest dry matter production when the planting density was high and planting dates was early. When silage was made from either MET line or check hybrid the nutritional values in terms of crude protein, crude fat, fiber, and ash contents of the MET line were similar to those of the check hybrid. But the TON of the MET line was higher than that of the check hybrid, while the OCP of the MET line lower than that of the check hybrid. Amino acid contents of the MET line were also comparable to those of the check hybrid, while lysine content of the MET line was 10% higher than that of the check hybrid.
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