Transglutaminase-catalyzed incorporation of lysine oligomers into casein

Bercovici, Daniel; Gaertner, Hubert; Puigserver, Antoine

doi:10.1021/jf00075a004

Cited by 18 publications

(7 citation statements)

References 22 publications

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“…Guinea-pig liver transglutaminase was obtained and purified to homogeneity as described by Brookhart et al [9], slightly modified by Bercovici et al [10].…”

Section: Purification Of Transglutaminasementioning

confidence: 99%

Molecular mechanisms of the irreversible thermal denaturation of guinea-pig liver transglutaminase

Nury

Meunier

1990

Biochemical Journal

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When transglutaminase is heated at temperatures above 40 degrees C, it loses its activity according to a two-step mechanism [Nury, Meunier & Mouranche (1989) Eur. J. Biochem. 180, 161-166]: N----X(TD)----D However, the nature of the molecular events responsible for the irreversible denaturation is still unknown. Investigation of the effects of dithiothreitol and 5,5'-dithiobis-2-nitrobenzoate on the kinetics of inactivation, titrations of ammonia released by deamidation and of thiol groups on the native and denatured enzymes and SDS/PAGE rule out the involvement of covalent processes during the denaturation of transglutaminase at 55 degrees C and pH 7. Of the two possible kinds of non-covalent events, i.e. unfolding of the polypeptide chain and aggregation of enzyme molecules, we show that both occur, though only the former process is responsible for the denaturation. The latter process, aggregation, follows the unfolding of the molecules.

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“…Guinea-pig liver transglutaminase was obtained and purified to homogeneity as described by Brookhart et al [9], slightly modified by Bercovici et al [10].…”

Section: Purification Of Transglutaminasementioning

confidence: 99%

Molecular mechanisms of the irreversible thermal denaturation of guinea-pig liver transglutaminase

Nury

Meunier

1990

Biochemical Journal

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“…Transglutaminase or protein-glutamine y-glutamyltransferase (EC 2.3.2.13); glutamate dehydrogenase or L-glutamate: NAD oxidoreductase (EC 1.4.1.3). modified by Bercovici et al [14]. Livers in all cases were perfused.…”

Section: Purification Of ' Transglutaminasementioning

confidence: 99%

The kinetics of the thermal deactivation of transglutaminase from guinea‐pig liver

Nury

Meunier

Mouranche

1989

European Journal of Biochemistry

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By incubating native (N) transglutaminase from guinea-pig liver at various temperatures and assaying it at 2572, two steps in the irreversible deactivation process to the denatured form (D) have been found. The fitting of the data to the equations of two possible models (the two-steps model and the two-isoenzymes model) is only compatible with the first one (N ---* X + D). It is shown that the structure of the active intermediate, X, depends on the deactivation temperature and on the thermal history of the enzyme. This may mean that transglutaminase exists in a large number of microstates. Surprisingly, the activation energy of deactivation is lower than that of activity (36.6 & 3.4 against 47.2 2.2 kJ . mol-I). By deactivating transglutaminase at a constant temperature (55°C) and assaying it at variable temperatures, the activation energy of the intermediate, (X55), has been determined to be 40.2 _+ 5 kJ . mol-l, of the same order of magnitude as the native form.Among several agents assayed, only Ca2+ had a positive effect on the thermal stability of this enzyme. At 40°C, transglutaminase was quite stable in the presence of Ca2+ (in its absence, the half-life was 65 min) and at 45 "C, its thermostability had been considerably increased, the half-life being raised from 47 min to 275 min.Transglutaminase is a Ca2 +-dependent enzyme responsible for post-translational modifications of proteins. It allows the cross-linking of proteins like fibrin, collagen and uteroglobulin [l -51. It is a tool for in vitro modifications ofproteins and especially for making gels [6 -81. Although the kinetics of the reversible thermal deactivation of proteins (and especially enzymes) has been extensively studied [9], the kinetics of the irreversible process has been the object of only a few papers [lo -121. To increase our knowledge in this basic area, we have carried out such a study on the transglutaminase from guinea-pig liver. This basic study may also be of some interest on potential uses of this enzyme. Knowledge of the kinetics of its irreversible deactivation may be essential for the development of rational approaches to enzyme stabilization, e. g. enhancement of enzyme thermostability by such agents as CaZ + . Therefore, the aim of this paper was to study the effect of temperature on the activity and on the stability of transglutaminase from guinea-pig liver and the role of Ca2+ in the enhancement of its thermostability. MATERIALS AND METHODS Purification of ' transglutaminaseEnzyme was extracted and purified from guinea-pig liver according to the method of Brookhart et al. [13], slightlyCorrespondence to J

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“…Since the 1980s, to improve the quality and nutritive value of food, research has focused on the potential use of TGs in food processing, exploiting their ability to catalyse intermolecular isopeptide bonds and polymerise proteins [ 115 , 116 , 117 ]. Hence, different groups begun investigating the best substrates for TGs activity in this area and many were identified, such as dairy proteins (e.g., caseins and whey proteins) [ 118 , 119 , 120 ], soybean globulins [ 121 ], wheat (gluten) [ 122 , 123 ], myosins [ 124 , 125 ], egg [ 126 ], and seafood proteins [ 127 , 128 ].…”

Section: Tgs Crosslinking Activity In Biotechnological Applicationmentioning

confidence: 99%

Biocatalysis by Transglutaminases: A Review of Biotechnological Applications

et al. 2018

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The biocatalytic activity of transglutaminases (TGs) leads to the synthesis of new covalent isopeptide bonds (crosslinks) between peptide-bound glutamine and lysine residues, but also the transamidation of primary amines to glutamine residues, which ultimately can result into protein polymerisation. Operating with a cysteine/histidine/aspartic acid (Cys/His/Asp) catalytic triad, TGs induce the post-translational modification of proteins at both physiological and pathological conditions (e.g., accumulation of matrices in tissue fibrosis). Because of the disparate biotechnological applications, this large family of protein-remodelling enzymes have stimulated an escalation of interest. In the past 50 years, both mammalian and microbial TGs polymerising activity has been exploited in the food industry for the improvement of aliments’ quality, texture, and nutritive value, other than to enhance the food appearance and increased marketability. At the same time, the ability of TGs to crosslink extracellular matrix proteins, like collagen, as well as synthetic biopolymers, has led to multiple applications in biomedicine, such as the production of biocompatible scaffolds and hydrogels for tissue engineering and drug delivery, or DNA-protein bio-conjugation and antibody functionalisation. Here, we summarise the most recent advances in the field, focusing on the utilisation of TGs-mediated protein multimerisation in biotechnological and bioengineering applications.

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Transglutaminase-catalyzed incorporation of lysine oligomers into casein

Cited by 18 publications

References 22 publications

Molecular mechanisms of the irreversible thermal denaturation of guinea-pig liver transglutaminase

Molecular mechanisms of the irreversible thermal denaturation of guinea-pig liver transglutaminase

The kinetics of the thermal deactivation of transglutaminase from guinea‐pig liver

Biocatalysis by Transglutaminases: A Review of Biotechnological Applications

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