The structure of glutaraldehyde in aqueous solution determined by ultraviolet absorption and light scattering

Kawahara, Jun‐ichiro; Ohmori, Takao; Ohkubo, Teiji; Hattori, Shigeru; Kawamura, Mitsutaka

doi:10.1016/0003-2697(92)90178-a

Cited by 71 publications

(51 citation statements)

References 13 publications

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“…In tissues or proteins fixed with glutaraldehyde and osmium tetroxide, sodium metaperiodate treatment probably oxidizes the osmium additives completely and forms the 1, 2-dioles and cleaves the diols to aldehydes, thus inducing further degradation of the protein-glutaraldehyde co-polymers. The results of the present study support the hypothesis of Kawahara et al (1992;1997) concerning the mechanisms of fixation by glutaraldehyde.…”

Section: Discussionsupporting

confidence: 90%

“…In particular, several investigators have stated that α, β-unsaturated oligomers/polymers play a central role in the crosslink reaction of proteins by producing Schiff bases, as the crosslinking efficiency of glutaraldehyde is remarkably higher than that of other bi-functional, crosslinking reagents, and because glutaraldehyde forms extremely stable crosslinks against acid hydrolysis, with the crosslinks being stable in 6 N hydrochloric acid at 110C for 24 hr (Monsan et al 1975;Migneault et al 2004). In contrast, Kawahara et al (1992) demonstrated that the aldol condensation of glutaraldehyde monomers and protein crosslinking by Schiff bases should occur in parallel and should form a linear aldol-condensed glutaraldehyde oligomer with several Schiff base linkages branching off, as the glutaraldehyde solution showed no absorbance at 235 nm caused by the α, β-unsaturated bonds in the absence of amines (Fig. 6(b)).…”

Section: Discussionmentioning

confidence: 96%

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Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Yamashita

Okada

2014

J Histochem Cytochem.

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SummaryWe studied the effectiveness of heat-induced antigen retrieval (HIAR) in conventionally processed, epon-embedded specimens and the mechanisms of HIAR in the specimens. Frozen sections were first immunostained to examine the possibility of using HIAR for 18 antigens to avoid the effects of epoxy resin embedment. The antigenicity of 7 out of 18 antigens was retrieved with glutaraldehyde fixation followed by osmium tetroxide treatment whereas none were retrieved with glutaraldehyde fixation without post-osmication. Six antigens also exhibited positive immunostaining in semi-thin epon sections when the sections were deplasticized with sodium ethoxide followed by autoclaving. In the immunoelectron microscopy with the post-embedding method, positive reactions with fine ultrastructures were obtained using HIAR without deplasticization. These results suggested that osmium tetroxide binds to ethylene double bonds (which are introduced into protein crosslinks by glutaraldehyde) and forms an extremely stable resonance interaction with the Schiff bases, thus destabilizing the protein crosslinks. Heating also further degrades these crosslinks. The present study demonstrated that archival epon blocks can be useful resources for immunohistochemical studies for both light and electron microscopy. (J Histochem Cytochem 62:584-597, 2014) Keywords heat-induced antigen retrieval, osmicated and epon-embedded specimen, immunoelectron microscopy, mechanism of antigen retrieval

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 96%

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Yamashita

Okada

2014

J Histochem Cytochem.

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“…This conversion had been suggested to occur by several researchers (18, 20, 26), but its chemical, physiological and ecological consequences had not been studied in detail. As glutaraldehyde is well known as an potent alkylator, crosslinker, and denaturant of protein and is used to fix protein in histochemical studies (27)(28)(29)(30)(31), we hypothesized that the same reaction might occur in our system. To examine this possibility, we mixed 11 mM oleuropein with 5 units of ␤-glucosidase from sweet almond.…”

Section: Resultsmentioning

confidence: 99%

“…These studies reported that the adduct formed between glutaraldehyde and amine is irreversible under acid hydrolysis, whereas the adduct formed between simple aldehyde and amine is reversible (27). Interestingly, the molecular structure of glutaraldehyde in aqueous solution and the chemical mechanism of the irreversible protein-crosslinking are still unclear and remain under debate (27)(28)(29)(30)(31), although several possible mechanisms have been proposed (see ref. 27 for review).…”

Section: Discussionmentioning

confidence: 99%

Enzymatic activation of oleuropein: A protein crosslinker used as a chemical defense in the privet tree

Konno

Hirayama²,

Yasui³

et al. 1999

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

202

174

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Leaves of the privet tree, Ligustrum obtusifolium, contain a large amount of oleuropein, a phenolic secoiridoid glycoside, which is stably kept in a compartment separate from activating enzymes. When the leaf tissue is destroyed by herbivores, enzymes localized in organelles start to activate oleuropein into a very strong protein denaturant that has protein-crosslinking and lysine-decreasing activities. These activities are stronger than ever reported from plant systems and have adverse effects against herbivores by decreasing the nutritive value of dietary protein completely. We report here that strong oleuropein-specific ␤-glucosidase in organelles activates oleuropein by converting the secoiridoid glucoside moiety of oleuropein into a glutaraldehyde-like structure, which is also an ␣,␤-unsaturated aldehyde. Oleuropein activated by ␤-glucosidase had very strong proteindenaturing, protein-crosslinking, and lysine-alkylating activities that are very similar to, but stronger than, those of glutaraldehyde. Aucubin, another iridoid glycoside, had similar activities after ␤-glucosidase treatment. We also detected polyphenol oxidase activity in organelles that activate the dihydroxyphenolic moiety to have protein-crosslinking activities. These data suggest that the privet tree has developed an effective defense mechanism with oleuropein, a unique multivalent alkylator ideal as a protein-crosslinker. Our results that iridoid glycosides are precursors of alkylators may elucidate the chemical bases that underlie various bioactivities and ecological roles of iridoid glycosides.

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“…Furthermore, in anhydrous solvents the equilibrium between monomeric and polymerized GA possibly shifts to the latter, which produces water. 8 Some researchers made experiments in D20. 2 • 3 However, since deuterium exchange could occur at a -carbon,9 it might give erroneous results to compare the peak intensities of 1 H-NMR.…”

mentioning

confidence: 99%

Chemical Cross-Linking by Glutaraldehyde between Amino Groups: Its Mechanism and Effects

Kawahara

Ishikawa

Uchimaru

et al. 1997

Polymer Modification

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The structure of glutaraldehyde in aqueous solution determined by ultraviolet absorption and light scattering

Cited by 71 publications

References 13 publications

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Enzymatic activation of oleuropein: A protein crosslinker used as a chemical defense in the privet tree

Chemical Cross-Linking by Glutaraldehyde between Amino Groups: Its Mechanism and Effects

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