• BMB Reports
• /
• v.42 no.2
• /
• pp.113-118
• /
• 2009

Despite the growing body of evidence suggesting a role for MsrA in antioxidant defense, little is currently known regarding the function of MsrB in cellular protection against oxidative stress. In this study, we overexpressed the mammalian MsrB and MsrA genes in Saccharomyces cerevisiae and assessed their subcellular localization and antioxidant functions. We found that the mitochondrial MsrB3 protein (MsrB3B) was localized to the cytosol, but not to the mitochondria, of the yeast cells. The mitochondrial MsrB2 protein was detected in the mitochondria and, to a lesser extent, the cytosol of the yeast cells. In this study, we report the first evidence that MsrB3 overexpression in yeast cells protected them against $H_2O_2$-mediated cell death. Additionally, MsrB2 overexpression also provided yeast cells with resistance to oxidative stress, as did MsrA overexpression. Our results show that mammalian MsrB and MsrA proteins perform crucial functions in protection against oxidative stress in lower eukaryotic yeast cells.

• BMB Reports
• /
• v.42 no.9
• /
• pp.580-585
• /
• 2009

Dimethyl sulfoxide (DMSO) is widely used in chemistry and biology as a solvent and as a cryoprotectant. It is also used as a pharmaceutical agent for the treatment of interstitial cystitis and rheumatoid arthritis. Previous reports described DMSO as being reduced by methionine-S-sulfoxide reductase (MsrA). However, little is known about the DMSO reduction capability of methionine-R-sulfoxide reductase (MsrB) or its effect on the catalysis of methionine sulfoxide reduction. We show that mammalian MsrB2 and MsrB3 were unable to reduce DMSO. This compound inhibited MsrB2 activity but did not inhibit MsrB3 activity. We further determined that DMSO functions as an inhibitor of MsrA and MsrB2 in the reduction of methionine sulfoxides via different inhibition mechanisms. DMSO competitively inhibited MsrA activity but acted as a non-competitive inhibitor of MsrB2 activity. Our study also demonstrated that DMSO inhibits in vivo methionine sulfoxide reduction in yeast and mammalian cells.

• Weed & Turfgrass Science
• /
• v.2 no.2
• /
• pp.159-163
• /
• 2013

The purpose of this study is to observe the growth habit and investigate a possibility of cultivating the GM rice (CaMsrB2-8) as a rice cultivar having drought resistance. Germination viability test showed that there was no significant difference between the drought-resistant GM(CaMsrB2-8) and non-GM (Ilmi) rice which was the parent variety at the GM rice. All the seeds of CaMsrB2-8 and Ilmi germinated after 6 days. Viviparous germination was not found in CaMsrB2-8 and Ilmi that was grown in greenhouse at $23{\pm}2^{\circ}C$ with water spraying for 40 days. Ratooning of CaMsrB2-8 and Ilmi was observed in 7-14 days and found uniform in field condition. CaMsrB2-8 seemed to grow faster than Ilmi. But CaMsrB2-8 and Ilmi were similar in 14-21 days. Both CaMsrB2-8 and Ilmi showed low seed shattering and more than 90% grains were ripened. All the seeds scattered in the paddy soil surface were not germinated after passing the winter. This study suggests that the drought-resistant GM rice was not significantly different with the parent variety of Ilmi in many agronomic characteristics such as wildness traits.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.1
• /
• pp.59-62
• /
• 2008

The peptide methionine sulfoxide reductases (Msrs) are enzymes that catalyze the reduction of methionine sulfoxide back to methionine. Because of two enantiomers of methionine sulfoxide (S and R forms), this reduction reaction is carried out by two structurally unrelated classes of enzymes, MsrA (E.C. 1.8.4.11) and MsrB (E.C. 1.8.4.12). Whereas MsrA has been well characterized structurally and functionally, little information on MsrB is available. The recombinant MsrB from Bacillus subtilis has been purified and crystallized by the hanging-drop vapor-diffusion method, and the functional and structural features of MsrB have been elucidated. The crystals belong to the trigonal space group P3, with unit-cell parameters a=b=136.096, $c=61.918{\AA}$, and diffracted to $2.5{\AA}$ resolution using a synchrotron-radiation source at Pohang Light Source. The asymmetric unit contains six subunits of MsrB with a crystal volume per protein mass $(V_M)\;of\;3.37{\AA}^3\;Da^{-1}$ and a solvent content of 63.5%.

• Progress in Superconductivity
• /
• v.9 no.1
• /
• pp.45-49
• /
• 2007

We have investigated the optimum combination of the environmental noise condition and type of SQUID pickup coil in order to obtain maximum signal-to-noise ratio (SNR). The measurement probe consists of 1st order gradiometer with pickup coils of 100 mm, 70 mm, and 50 mm baseline length, a 2nd order gradiometer with 50 mm baseline, and a magnetometer. The pickup coils are fabricated by winding Nb wire on a bobbin with 200 mm diameter. Noise and heart signal of a healthy male were measured by various SQUID sensors with different types of pickup coils in various magnetically shielded rooms (MSR), and compared to each other. The shielding factors were found to be 43 dB, 35 dB and 25 dB at 0.1 Hz for MSR-AS, MSR-BS, MSR-CS, respectively. White noises were $3.5\;fT/Hz^{1/2}$, $4.5\;fT/Hz^{1/2}$ and $3\;fT/Hz^{1/2}$ for the 1st order gradiometers, the 2nd order gradiometers, and magnetometer for all MSRs. SNR of the magnetometer was up to 56 dB in MSR-AS, while the 1st order axial gradiometer with 70 mm baseline length was up to 54 dB in MSR-BS. The 2nd order axial gradiometer with 50 mm baseline length of pickup coil was found to be up to 40 dB in MSR-CS.

• BMB Reports
• /
• v.44 no.4
• /
• pp.256-261
• /
• 2011

PEP-1 peptide has been used for transduction of native protein into mammalian cells. This work describes the findings that the fusion of PEP-1 to target proteins led to protein truncation likely in a non-protein-specific manner. Approximately 75% of PEP-1-MsrA fusion protein was truncated in the N-terminal region of MsrA between Lys-27 and Val-28 during expression in Escherichia coli and purification. This large protein truncation was also observed in another PEP-1 fused protein, PEP-1-MsrB2, in the N-terminal region of MsrB2. The full-length PEP-1-MsrA protein was rapidly transduced into keratinocyte cells within 15 min. The transduced PEP-1-MsrA was functionally active and could protect skin cells against oxidative stress- and ultraviolet radiation-induced cell death. Collectively, our data demonstrated the protective roles of MsrA in skin cells and, moreover, may raise a concern of protein truncation caused by fusion of PEP-1 about the general use of this peptide for protein transduction.

• KSBB Journal
• /
• v.16 no.4
• /
• pp.415-419
• /
• 2001

To produce staphylokinase (SAK) in B. subtilis WB700, media optimization was carried out and the operation of batch and fed-batch fermentation were compared. Tryptone is a good nitrogen source and its optimum concentration in modified super rich(MSR) media is 15 g/L. When glucose is used as a limiting carbon source in the MSR media, 5 g/L of an optimum glucose concentration was identified for the SAK production under the control of P43 promoter. As the expression of P43 promoter is controlled by the limitation of oxygen, the SAK production was controlled at the 30% DO level in the fed-batch fermentation. Unexpectedly, batch fermentation using MSR media showed 1.5 times higher yield of SAK than that of the fed-batch fermentation. The main cause of the results comes from not achieving higher cell concentration in the fed-batch fermentation and the optimum expression level of P43 promoter under oxygen or nutrient limitations. We could not achieve the increase in cell concentration by any means in batch culture as well as fed-batch culture. The highest yield in the batch culture was 2880 units of SAK activity and 455 mg/L of secreted SAK.

• Current Research on Agriculture and Life Sciences
• /
• v.31 no.2
• /
• pp.124-130
• /
• 2013

The significance of environment change and genetic safety has been recently recognized by many genetically modified (GM) plants. This study was to evaluate the safety of drought-tolerant rice and to identify the environment variance. The GM rice of drought-tolerant rice and four check cultivars were analyzed the data on agronomic characters and principal component in large-GM crop field. There was no significant difference in agronomic characters between the drought-tolerant rice and donor plant, 'Ilmi'. Grain yield showed the standard deviation of the difference, did not significant statistically. Related to grain characters, grain appearance were similar to the drought-tolerant rice and donor plant, 'Ilmi'. In Chemical characters, brown rice of the drought-tolerant rice and a donor plant, 'Ilmi' did difference in starch and protein, however, was similar as 'Ilpum'. These results indicated that drought-tolerant rice may perform to detect genetic safety in GM plants progeny.

• BMB Reports
• /
• v.43 no.9
• /
• pp.622-628
• /
• 2010

Dimethyl sulfoxide (DMSO) can be reduced to dimethyl sulfide by MsrA, which stereospecifically catalyzes the reduction of methionine-S-sulfoxide to methionine. Our previous study showed that DMSO can competitively inhibit methionine sulfoxide reduction ability of yeast and mammalian MsrA in both in vitro and in vivo, and also act as a non-competitive inhibitor for mammalian MsrB2, specific for the reduction of methionine-R-sulfoxide, with lower inhibition effects. The present study investigated the effects of DMSO on the physiological antioxidant functions of methionine sulfoxide reductases. DMSO elevated hydrogen peroxide-mediated Saccharomyces cerevisiae cell death, whereas it protected human SK-Hep1 cells against oxidative stress. DMSO reduced the protein-carbonyl content in yeast cells in normal conditions, but markedly increased protein-carbonyl accumulation under oxidative stress. Using Msr deletion mutant yeast cells, we demonstrated the DMSO's selective inhibition of the antioxidant function of MsrA in S. cerevisiae, resulting in an increase in oxidative stress-induced cytotoxicity.

• BMB Reports
• /
• v.44 no.10
• /
• pp.669-673
• /
• 2011

Human methionine sulfoxide reductase B3A (hMsrB3A) is an endoplasmic reticulum (ER) reductase that catalyzes the stereospecific reduction of methionine-R-sulfoxide to methionine in proteins. In this work, we identified an antimicrobial peptide from hMsrB3A protein. The N-terminal ER-targeting signal peptide (amino acids 1-31) conferred an antimicrobial effect in Escherichia coli cells. Sequence and structural analyses showed that the overall positively charged ER signal peptide had an Argand Pro-rich region and a potential hydrophobic ${\alpha}$-helical segment that contains 4 cysteine residues. The potential ${\alpha}$-helical region was essential for the antimicrobial activity within E. coli cells. A synthetic peptide, comprised of 2-26 amino acids of the signal peptide, was effective at killing Gram-negative E. coli, Klebsiella pneumoniae, and Salmonella paratyphi, but had no bactericidal activity against Gram-positive Staphylococcus aureus.