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261.
Jinhee Lee Ahyun Son Paul Kim Soon Bin Kwon Ji Eun Yu Gyoonhee Han Baik L. Seong 《Biotechnology and bioengineering》2019,116(3):i-i
Transglutaminase (TGase) induces the cross-linking of proteins by catalyzing an acyl transfer reaction. TGase is a zymogen, activated by the removal of its pro-region. Because the pro-region is crucial for folding and inhibition of the TGase activity, the recombinant expression of the mature TGase (mTGase) without the pro-region, usually results in inactive inclusion bodies or low protein yield. Here, Streptomyces netropsis TGase was fused with Escherichia coli lysyl-tRNA synthetase (LysRS), as a module with chaperoning activity in an RNA dependent manner (chaperna). The TGase activity from purified fusion protein induced via the removal of LysRS by tev protease in vitro. Moreover, active mTGase was produced in E. coli via an intracellular cleavage system, wherein LysRS-mTGase was cleaved by the coexpressed tev protease in vivo. The results suggest that LysRS essentially mimics pro-region, which exerts a dual function—folding of TGase into active conformation and keeping it as dormant state—in an RNA-dependent manner. Thus, trans-acting RNAs, prompt the cis-acting chaperone function of LysRS, while being mechanistically similar to the intramolecular chaperone function of the pro-region. These results could be implemented and extended for the folding of “difficult-to-express” recombinant proteins, by harnessing the chaperna function. 相似文献
262.
A facile approach for the production of a reusable immobilized recombinant Escherichia coli biotin ligase (BirA) onto amine-modified magnetic microspheres (MMS) via covalent cross-linking catalyzed using microbial transglutaminase (MTG) was proposed in this study. The site-specifically immobilized BirA exhibited approximately 95% of enzymatic activity of the free BirA, and without a significant loss in intrinsic activity after 10 rounds of recycling (P > 0.05). In addition, the immobilized BirA can be easily recovered from the solution via a simple magnetic separation. Thus, the immobilized BirA may be of general use for in vitro biotinylation in an efficient and economical manner. 相似文献
263.
Cellular transglutaminase has affinity for extracellular matrix 总被引:4,自引:0,他引:4
Herbert F. Upchurch Eugene Conway M. K. Patterson Jr. Paul J. Birckbichler Merle D. Maxwell 《In vitro cellular & developmental biology. Plant》1987,23(11):795-800
Summary Cellular transglutaminase (TGase) was demonstrated as an intracellular enzyme by immunofluorescence in WI-38 cells. Following
cell membrane perturbation by Triton X-100 treatment, TGase was bound to the extracellular matrix and was found to coexist
with fibronectin as visualized by immunofluorescence microscopy. The binding of TGase to the cell matrix was blocked by anti-fibronectin
antibody. Exogenous sources of soluble TGase were transferred to the extracellular matrix of an untreated or methanol fixed
cell. The experimental data indicated that “particulate bound” TGase is a consequence of soluble TGase binding to the extracellular
matrix following cell rupture.
Editor's statement This report suggest that “particulate bound” transglutaminase may be a consequence of affinity of soluble
enzyme for specific molecules in extracellular matrix and opens up a means to characterize transglutaminase binding sites
in the matrix. 相似文献
264.
Rat pancreatic islet homogenates catalyze the incorporation of [2,5–3-H]histamine into endogenous proteins recovered in both the stacking gel and a Mr 84000 protein separated by polyacrylamide electrophoresis. The labelling of these proteins represents a Ca2+-dependent process inhibited by glycine methylester, but not sarcosine methylester, and enhanced after preincubation of the islets at a high concentration of D-glucose. Although transglutaminase activity is found in both soluble and particlate subcelluler fractions, the endogenous transglutaminase substrates were located mainly in paarticulate, possibly membrane-associated, material. 相似文献
265.
Thomas M. Jeitner John T. Pinto† Boris F. Krasnikov† Mark Horswill‡ Arthur J. L. Cooper† 《Journal of neurochemistry》2009,109(S1):160-166
Transglutaminases (TGs) are Ca2+ -dependent enzymes that catalyze a variety of modifications of glutaminyl (Q) residues. In the brain, these modifications include the covalent attachment of a number of amine-bearing compounds, including lysyl (K) residues and polyamines, which serve to either regulate enzyme activity or attach the TG substrates to biological matrices. Aberrant TG activity is thought to contribute to Alzheimer disease, Parkinson disease, Huntington disease, and supranuclear palsy. Strategies designed to interfere with TG activity have some benefit in animal models of Huntington and Parkinson diseases. The following review summarizes the involvement of TGs in neurodegenerative diseases and discusses the possible use of selective inhibitors as therapeutic agents in these diseases. 相似文献
266.
Li Cui Guocheng Du Qiang Wang Ping Wang Xuerong Fan 《Biocatalysis and Biotransformation》2013,31(5):399-404
The dyeing properties of three natural dyes – curcumin, gardenia yellow and lac dye – on wool fabric after treatment with microbial transglutaminase (MTGase) have been investigated. After 120 min of MTGase treatment, compared with the fabric only pretreated with chemical and protease, the colour strength of curcumin, gardenia yellow and lac dye increased from 8±0.13, 7.5±0.10 and 22±0.12 to about 12.8±0.20, 11.7±0.20 and 27.0±0.41, respectively. The values of wash fastness for dyed wool fabrics increased from 2 to 4 after MTGase treatment, but the light fastness was not obviously improved. By comparing with mordant dyeing, although the colour strength was poorer, MTGase after-treatment did not cause colour shade changes during dyeing and the wash fastness of dyed wool fabric was similar to that of the pre-mordanted samples. 相似文献
267.
Crude lipase from Rhizopus sp. was moderately stable against heat treatment at 45 °C. However, after incubation for 1 h at 25 °C with Streptoverticillium transglutaminase (MTG), the half-life of crude lipase in the heat treatment was increased more than 10-fold compared to that of untreated one. The result can be ascribed by the MTGase-mediated crosslinking of contaminating proteins that affect the apparent thermostability of lipase in the crude sample. 相似文献
268.
Quantitative and rapid analysis of transglutaminase activity using protein arrays in mammalian cells
Mi-Hye Kwon Jae-Wan Jung Se-Hui Jung Jin-Young Park Young-Myeong Kim Kwon-Soo Ha 《Molecules and cells》2009,27(3):337-343
We developed a novel on-chip activity assay using protein arrays for quantitative and rapid analysis of transglutami-nase
activity in mammalian cells. Transglutaminases are a family of Ca2+-dependent enzymes involved in cell regulation as well as human diseases such as neurodegenerative disorders, inflammatory
diseases and tumor progression. We fabricated the protein arrays by immobilizing N,N′-dimethylcasein (a substrate) on the
amine surface of the arrays. We initiated transamidating reaction on the protein arrays and determined the transglutaminase
activity by analyzing the fluorescence intensity of biotinylated casein. The on-chip transglutaminase activity assay was proved
to be much more sensitive than the [3H]putrescine-incorporation assay. We successfully applied the on-chip assay to a rapid and quantitative analysis of the transgluta-minase
activity in all-trans retinoic acid-treated NIH 3T3 and SH-SY5Y cells. In addition, the on-chip transglutaminase activity
assay was sufficiently sensitive to determine the transglutaminase activity in eleven mammalian cell lines. Thus, this novel
on-chip transglutaminase activity assay was confirmed to be a sensitive and high-throughput approach to investigating the
roles of transglutaminase in cellular signaling, and, moreover, it is likely to have a strong potential for monitoring human
diseases.
These authors contributed equally to this work. 相似文献
269.
Kimikazu Fujita Masato Ando Masamitsu Yamauchi Yutaka Nagata Masao Honda 《Neurochemical research》1995,20(10):1195-1201
We measured the activity of transglutaminase (TG), a Ca2+-dependent enzyme and a biochemical marker of cell degeneration, in the adult rat spinal cord after unilateral occlusion of a branch of the dorsal spinal artery, and compared it to the enzyme activity in the tissue on the contralateral side without ischemic damage. The affected half of the spinal cord showed a significant rise in intrinsic (endogenous) TG activity one day after ischemic insult while no apparent morphological changes were observed in the tissue. However, the enzymic activity on the affected side gradually decreased to reach the level in the non-affected tissue, accompanying severe degeneration of neuronal cells at 7 days after the surgery, then it declined to nearly half the level in the intact tissue 30 days after the operation. We also determined the TG activity in transverse sections of the human spinal cord obtained at autopsy from 5 amyotrophic lateral sclerosis (ALS) and 9 non-ALS patients. TG activity in thoracic and lumbar cords was markedly low in ALS patients not only in ventral and lateral regions but also in the dorsal portion. These findings imply that the reduced TG activity in the ALS spinal cord is one of the characteristic features of the disease reflecting exhaustion of the enzyme in the tissue resulting from degeneration of the spinal neurons through cross-linkage of soluble intraneuronal cytoplasmic proteins. 相似文献