Dansylcadaverine specific staining for transamidating enzymes

Lóránd, L.; Siefring, Gerald E.; Tong, Yam Shun; Bruner-Lorand, Joyce; Gray, A.

doi:10.1016/s0003-2697(79)80178-5

Cited by 52 publications

(32 citation statements)

References 24 publications

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“…1). Thus, brain TGase has electrophoretic mobility similar to the mobilities of other mammalian TGases (9,20,21,28). No fluorescent spot appeared in the absence of exogenous Ca2+ and in the presence of 5 mM EDTA, indicating the calcium dependence ofbrain TGase.…”

Section: Resultsmentioning

confidence: 77%

“…For initial detection of TGase in human brain, low-speed (1,000 x g) supernates of cortical homogenates were electrophoresed on dimethylcasein-containing agarose gels and allowed to react with dansylcadaverine (20). A fluorescent spot indicating the presence of TGase activity was readily apparent.…”

Section: Resultsmentioning

confidence: 99%

“…Thereafter, homogenates were spun at 200,000 X g, and the highspeed supernate was used as the source of enzyme in all subsequent studies. TGase was detected by the gel assay of Lorand et al (20), in which the fluorescent amine donor, dansylcadaverine, is covalently bound to an exogenous protein substrate, N,N'-dimethylcasein, the glutaminyl side chains ofwhich serve as the amine acceptors. Aliquots (20-50 p1) of brain supernates (6-7 mg of protein per ml) as well as the supernates of washed human erythrocyte lysates were electrophoresed in 1% agarose gels containing 0.5% dimethylcasein at 3 mA/cm for 4 hr at 40C.…”

mentioning

confidence: 99%

“…After gel fixation and washing, TGase activity was visualized under UV light as a fluorescent spot where dansylcadaverine had been bound by the enzyme in the gel to the dimethylcasein substrate. The technique is both sensitive and specific for transamidases of the endo-y-glutamine:Elysine transferase type (20,21).…”

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confidence: 99%

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Brain transglutaminase: in vitro crosslinking of human neurofilament proteins into insoluble polymers.

Selkoe¹,

Abraham²,

Ihara³

1982

Proc. Natl. Acad. Sci. U.S.A.

200

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The accumulation in aged human neurons of insoluble, high molecular weight filamentous polymers apparently linked by nondisulfide covalent bonds led us to examine human brain for the presence of transglutaminase (EC 2.3.2.13) and endogenous protein substrates for this crosslinking enzyme. We demonstrate the presence in brain of a transamidating enzyme that can covalently crosslink brain proteins into insoluble polymers in vitro by forming 'y-glutamyl-e-lysine intermolecular bridges.Brain transglutaminase is Ca2" dependent, has an electrophoretic mobility similar to that of erythrocyte transglutaminase, and is active in human postmortem brain from aged normal individuals and patients with Alzheimer disease (senile dementia). Brain neurofilament fractions incubated in the presence of transglutaminase, Ca2+, and the fluorescent amine dansylcadaverine form a fluorescent, nondisulfide-bonded insoluble polymer; this process is associated with a decrease in the amount of soluble neurofilament polypeptides in the preparation. Electron microscopy of the polymeric material reveals an extensive network ofconnecting fiaments, which can be immunostained with various neurofilament antisera. Cystamine, an inhibitor of transglutaminase, prevents the neurofilament crosslinking. Glial filaments and myelin basic protein can also serve as substrates of brain transglutaminase in vitro. Although Alzheimer disease-type paired helical filaments are not formed under the specific in vitro conditions employed, the data suggest one possible mechanism for the covalent crosslinking of filaments into insoluble polymers during human neuronal aging.

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Section: Resultsmentioning

confidence: 77%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Brain transglutaminase: in vitro crosslinking of human neurofilament proteins into insoluble polymers.

Selkoe¹,

Abraham²,

Ihara³

1982

Proc. Natl. Acad. Sci. U.S.A.

200

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“…la; numbers above closed circles). On Western blots of liver samples two transglutaminase bands could be visualized with small molecular mass difference representing isoforms of tissue transglutaminase in rodents [23]. The intensity of both bands was increased when the liver samples undergoing apoptosis were analysed ( fig.2b).…”

Section: Resultsmentioning

confidence: 99%

Induction and activation of tissue transglutaminase during programmed cell death

1987

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During the involution of lead nitrate-induced hyperplasia in rat liver a significant increase of transglutaminase activity, enzyme concentration, transglutaminase messenger RNA and protein-bound e-@-glutamyl)lysine (product of transglutaminase action) coincided with programmed death (apoptosis) of hepatocytes. Immunohistochemical examination showed the appearance of transglutaminase in apoptotic hepatocytes. An increased transglutaminase level was also detected during glucocorticoid-induced apoptosis of rat thymocytes.

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Presence of a proteinase inhibitor in the vitelline envelope and its role in eggs of the fishTribolodon hakonensis

Kudo

1998

J. Exp. Zool.

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Dansylcadaverine specific staining for transamidating enzymes

Cited by 52 publications

References 24 publications

Brain transglutaminase: in vitro crosslinking of human neurofilament proteins into insoluble polymers.

Brain transglutaminase: in vitro crosslinking of human neurofilament proteins into insoluble polymers.

Induction and activation of tissue transglutaminase during programmed cell death

Presence of a proteinase inhibitor in the vitelline envelope and its role in eggs of the fishTribolodon hakonensis

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