Isolation and characterization of brain-specific transglutaminases from rat

Kwak, Sahng Jung; Kim, Soo Youl; Kim, Yong Sik; Song, Kye Yong; Kim, In Gyu; Park, Sang Chul

doi:10.1038/emm.1998.26

Cited by 13 publications

(4 citation statements)

References 35 publications

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“…Many previous studies have been used different types of ion-exchange chromatography (SP-Sepahrose, DEAE-cellulose, Q Sepharose and DEAE-Sepharose) for purification of TGase (Ha and Iuchi, 1997;Kumazawa et al, 1997). The current study was in agreement with Goldsmith and Martin, (1995) and Kwak et al, (1998). And also the present study for purification of TGase by gel filtration Sephadex G 100 was in agreement with Folk, (1970); Tokunaga and Iwanaga (1993) and El Hofy et al, (2014).…”

Section: Transglutaminase Purificationsupporting

confidence: 86%

Characterization of Transglutaminase Isolated from Rosemary Leaves, and Development of Chemical, Rheological and Sensory Properties of Kareish Cheese Made with Transglutaminase.

Darwish

Taher

2017

Journal of Food and Dairy Sciences

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Purification of transglutaminase (TGase) isolated from leaves of rosemary (Rosmarinus officinalis L) carried out by three steps including protein precipitation with using ammonium sulfate (40-80%), followed by elution into ion exchange column (DEAE-Sephadex A-50). The further purification of dialyzed fractions were applied on Sephadex G-100. The purified enzyme eluted from gel filtration column (Sephadex G-100) had 26.51 yield associated with increased about 6.3 in fold of purification. The optimum pH for rosemary TGase was 6.0 and the highest activity of this enzyme associated with incubated at 60°C for catalytic reaction of Z-Gln-Gly and hydroxylamine. The TGase was stable following heating up to 70 for 15 min, while a complete loss of TGase activity was associated with exposing to 90°C for 5 min. The exposure of TGase to salt concentrations ranging from 2-4% did not effect on its activity. However increased NaCl concentrations of 5-14% caused a decline in the activity. The effects of cross-linked by different concentrations of TGase (2.5, 5 and 10U/g protein) on chemical, sensorial and textural of Kareish during 15 days of storage period were studied. The Kareish cheese chemical composition presented to be affected by using different concentration of TGase. The moisture and yield were the highest values in cheese made by 10U/g protein of TGase, while pH was the lowest value in the same treatment, compare with other concentrations of TGase and control. Kareish cheese made by using 10 10U/g protein of TGase receive high scores in sensorial properties. Hardness, adhesiveness, gumminess and chewiness values were decreased significantly (P< 0.05) in Kareish cheese made with TGase than control.

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Section: Transglutaminase Purificationsupporting

confidence: 86%

Characterization of Transglutaminase Isolated from Rosemary Leaves, and Development of Chemical, Rheological and Sensory Properties of Kareish Cheese Made with Transglutaminase.

Darwish

Taher

2017

Journal of Food and Dairy Sciences

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“…Ion-exchange chromatography, such as DEAE-Sepharose, DEAE--cellulose, QSepharose, and SP-Sepharose, have been used to purify other TGase [Tokunaga and Iwanaga 1993, Ha and Iuchi 1997, Kumazawa et al 1997. The elution profi le from DEAE-Sephadex A-50 chromatography of rosemary TGase was similar from other TGases such as human epidermal TGase was eluted at 0.2 M NaCl [Goldsmith and Martin 1995], and rat brain TGase was observed to be eluted at 0.28 M and 0.35 M of NaCl concentration [Kwak et al 1998]. The fi nal step of purifi cation was achieved *Based on 50 g rosemary.…”

Section: Purifi Cation Of Transglutaminasementioning

confidence: 99%

Isolation, purifi cation and characterisation of transglutaminase from rosemary (Rosmarinus officinalis L.) leaves

El-Hofi

Ismail

Nour

et al. 2014

Acta Sci Pol Technol Aliment

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Background. Rosemary (Rosmarinus offi cinalis L.) is a spice and medicinal herb widely used around the world of the natural antioxidants, and it has been widely accepted as one of the spices with the highest antioxidant activity. Transglutaminase (EC 2.3.2.13; TGase) is an enzyme capable of catalysing acyl transfer reactions by introducing covalent cross-links between proteins, as well as peptides and various primary amines. TGase activity in plants was fi rst observed in pea seedlings, and subsequently found in organs of both lower and higher plants. Recently, TGase has captured researchers' interest due to its attractive potential application in food industries. Therefore, the objectives of this study are isolation and purifi cation of TGase from new plant source rosemary (Rosmarinus offi cinalis L.) leaves at the laboratory scale. Moreover, investigation of the biochemical properties of the purifi ed TGase to provide a suitable TGase enzyme for food industry applications are in focus. Material and methods. Rosemary (Rosmarinus offi cinalis L.) leaves was used as a new plant source to TGase. The biochemical characteristics of the crude and purifi ed enzyme were determined. Results. Rosemary (Rosmarinus offi cinalis L.) TGase was purifi ed to homogeneity by successive three purifi cation steps including ammonium sulfate precipitatation, ion exchange chromatography on DEAE-Sephadex A-50 column and Size exclusion column chromatography on Sephadex G-100 column. Under experimental conditions, 20-30% of ammonium sulfate saturation in the enzyme solution had a high yield of enzyme activity could be obtained. The purifi ed enzyme from the Sephadex G-100 column had 21.35% yield with increased about 7.31 in purifi cation fold. Rosemary TGase exhibited optimum activity at pH 7.0 and 55°C for the catalytic reaction of hydroxylamine and Z-Gln-Gly. The purifi ed TGase almost maintained full activity after incubation for 15 min up to 60°C and it was completely inactivated at 85°C. The rosemary TGase was stimulated at 2-6 mM CaCl 2 concentrations while it lost about 5-20% from its activity by increasing CaCl 2 concentration. Sodium chloride (2-14%) shows no noticeable inhibition of the purifi ed TGase activity. Mg +2 , Ba +2 were acivited by the purifi ed TGase while it was strongly inhibited by Fe +2 , moderately by Cu +2 and Mn +2 . Conclusion. This paper reports on the purifi cation and characterisation of TGase from newly isolated plant, rosemary (Rosmarinus offi cinalis L.) leaves. Finding results of the TGase properties make this enzyme a good candidate for application in the food industry. However, additional work is required to increase activity yield during extraction and purifi cation for commercial scale of TGase from this plant.

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“…The theory can also explain why pathology is restricted to certain brain regions, despite wide-spread expression of mutated protein throughout the body in the various (CAG) n /Q n expansion diseases. Apparently, the rat brain contains at least two unique forms of tTGase [Kwak et al, 1998]. Moreover, recent work has shown that at least three transglutaminases, including tTGase, are present in the human brain .…”

Section: Implications For Neurodegenerative Diseasesmentioning

confidence: 99%

Lysine-Rich Histone (H1) Is a Lysyl Substrate of Tissue Transglutaminase: Possible Involvement of Transglutaminase in the Formation of Nuclear Aggregates in (CAG)<sub>n</sub>/Q<sub>n</sub> Expansion Diseases

Cooper

Wang²,

Pasternack³

et al. 2000

Dev Neurosci

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Histone H1, which contains about 27% lysine, is an excellent lysyl donor substrate of Ca²⁺-activated guinea pig liver tissue transglutaminase as judged by rapid fluorescence enhancement in the presence of the glutaminyl-donor substrate 1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5′N′N′-dimethylaminonaphthalenesulfonyl) diamidopentane. Sodium dodecyl sulfate gel electrophoresis of a 30-min reaction mixture revealed the presence of fluorescent high-M_r aggregates, which are also formed when histone H1 is incubated solely with activated tissue transglutaminase. Aggregate formation is even more pronounced when histone H1 is incubated with activated tissue transglutaminase and dimethylcasein (glutaminyl donor only). The findings suggest not only that histone H1 is an especially good lysyl substrate of tissue transglutaminase, but that it is also a glutaminyl substrate. Histone H1 is a good lysyl substrate of transglutaminase purified from Streptoverticillium mobaraense, suggesting that the ability of histone H1 to act as a transglutaminase lysyl substrate is widespread. In agreement with previous studies, it was found that human β-endorphin is a moderately good substrate of tissue transglutaminase. At least 8 neurodegenerative diseases, including Huntington’s disease, are caused by (CAG)_n expansions in the genome and by an expansion of the corresponding polyglutamine domain within the expressed, mutated protein. Polyglutamine domains are excellent substrates of liver and brain transglutaminases. A hallmark of many of the (CAG)_n/polyglutamine expansion diseases is the presence of polyglutamine-containing aggregates within the cytosol and nuclei of affected neurons. Transglutaminase activity occurs in both of these compartments in human brain. In future studies, it will be important to determine whether transglutaminases play a role in (1) cross-linking of histone H1 to glutaminyl donors (including polyglutamine domains) in nuclear chromatin, (2) the formation of nuclear aggregates in (CAG)_n/polyglutamine expansion diseases, (3) DNA laddering and cell death in neurodegenerative diseases and (4) depletion of neuropeptides in vulnerable regions of Huntington’s disease brain.

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Isolation and characterization of brain-specific transglutaminases from rat

Cited by 13 publications

References 35 publications

Characterization of Transglutaminase Isolated from Rosemary Leaves, and Development of Chemical, Rheological and Sensory Properties of Kareish Cheese Made with Transglutaminase.

Characterization of Transglutaminase Isolated from Rosemary Leaves, and Development of Chemical, Rheological and Sensory Properties of Kareish Cheese Made with Transglutaminase.

Isolation, purifi cation and characterisation of transglutaminase from rosemary (Rosmarinus officinalis L.) leaves

Lysine-Rich Histone (H1) Is a Lysyl Substrate of Tissue Transglutaminase: Possible Involvement of Transglutaminase in the Formation of Nuclear Aggregates in (CAG)<sub>n</sub>/Q<sub>n</sub> Expansion Diseases

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