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A novel gene, sps2, detected in mouse embryo at the early stages of development has been identified as an analog of the E. coli selenophosphate synthetase gene. Unlike the E. coli enzyme, the presence of selenocysteine in the mouse enzyme is indicated by a TGA codon in the open reading frame of the cDNA. Using an N-FLAG monoclonal antibody, it was shown that the full length N-FLAG-sps2 gene product was expressed in COS-7 cells. To investigate the biological activity of the sps2 gene product in vivo, the mutated sps2 gene, which contains cysteine in the place of the TGA encoded selenocysteine in the wild type, was expressed in the E. coli selD deficient mutant, MB08. Like the E. coli wild type selD gene, the mutant sps2 gene complemented the selD mutation. However, replacement of Cys with either Ala, Ser, or Thr resulted in a loss of ability to complement the selD mutation. The SPS2-CYS protein expressed in E. coli was purified and its catalytic activity was determined. The Km value for ATP was 0.75 mM and Vmax was 9.23 nmole/min/mg protein. These results confirm that the mouse embryonic sps2 gene encodes an eukaryotic selenophosphate synthetase, and that availability of selenophosphate as a selenium donor compound is widespread. 相似文献
3.
ComC is required for the processing and translocation of ComGC, a pilin-like competence protein of Bacillus subtilis 总被引:9,自引:4,他引:5
ComGC is a cell surface-localized protein required for DNA binding during transformation in Bacillus subtilis. It resembles type IV prepilins in its N-terminal domain, particularly in the amino acid sequence surrounding the processing cleavage sites of these proteins. ComC is another protein required for DNA binding, which resembles the processing proteases that cleave type IV prepilins. We show here that ComGC is processed in competent cells and that this processing requires ComC. We also demonstrate that the PilD protein of Neisseria gonorrhoeae, a ComC homologue, can process ComGC in Escherichia coli, and that the ComC protein itself is the only B. subtilis protein needed to accomplish cleavage of ComGC in the latter organism. Based on NaOH-solubility studies, we have shown that in the absence of ComC, but in the presence of all other competence proteins, B. subtilis is incapable of correctly translocating ComGC to the outer face of the cell membrane. Finally, we show that ComGC can be cross-linked to yield a form with higher molecular mass, possibly a dimer, and present evidence suggesting that formation of the higher mass complex takes place in the membrane, prior to translocation. Formation of this complex does not require ComC or any of the comG products, other than ComGC itself. 相似文献
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Surface Plasmon Resonance Imaging-Based Protein Array Chip System for Monitoring a Hexahistidine-Tagged Protein during Expression and Purification 下载免费PDF全文
Hyeon-Su Ro Sun Ok Jung Byung Hoon Kho Hyung Pyo Hong Jae Sung Lee Yong-Beom Shin Min Gon Kim Bong Hyun Chung 《Applied microbiology》2005,71(2):1089-1092
A surface plasmon resonance imaging-based Ni2+-iminodiacetic acid-coated gold chip system was developed to enable specific detection of a hexahistidine-tagged recombinant protein in crude extracts or in column chromatography fractions. This system is especially advantageous for high-throughput analysis of multiple proteins. 相似文献
6.
We present a rapid and high-throughput human serum N-glycan preparation technology using 96-well plate-based procedures. The released N-glycans from polyvinylidene fluoride (PVDF) membrane filter plate are subsequently loaded to porous graphitic carbon (PGC) containing a 96-well plate to remove salts and other contaminants without sacrificing accuracy or reproducibility. This robust glycan preparation technology is applied to ovarian cancer diagnosis using 5 μl of patient serum. 相似文献
7.
Chung Ill-Min Rekha Kaliyaperumal Rajakumar Govindasamy Thiruvengadam Muthu 《Bioprocess and biosystems engineering》2018,41(11):1665-1677
Bioprocess and Biosystems Engineering - Glucosinolates (GSLs) and phenolic compounds (PCs) are biologically active and involved in the defense reaction of plants; these compounds have a beneficial... 相似文献
8.
Ong Jia Sin Taylor Todd D. Yong Cheng Chung Khoo Boon Yin Sasidharan Sreenivasan Choi Sy Bing Ohno Hiroshi Liong Min Tze 《Probiotics and antimicrobial proteins》2020,12(1):125-137
Probiotics and Antimicrobial Proteins - This study aimed to elucidate the targets and mechanisms of anti-staphylococcal effects from bioactive metabolites produced by lactic acid bacteria. We aimed... 相似文献
9.
Spliceosome-targeted therapies trigger an antiviral immune response in triple-negative breast cancer
Elizabeth A. Bowling Jarey H. Wang Fade Gong William Wu Nicholas J. Neill Ik Sun Kim Siddhartha Tyagi Mayra Orellana Sarah J. Kurley Rocio Dominguez-Vidaña Hsiang-Ching Chung Tiffany Y.-T. Hsu Julien Dubrulle Alexander B. Saltzman Heyuan Li Jitendra K. Meena Gino M. Canlas Srinivas Chamakuri Thomas F. Westbrook 《Cell》2021,184(2):384-403.e21
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