Determination of Zinc in Biological Materials by Atomic Absorption Spectrometry.

Fuwa, Keiichirō; Pulido, Pablo; McKay, Roxane; Vallée, Bert L.

doi:10.1021/ac60219a009

Cited by 148 publications

(65 citation statements)

References 19 publications

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“…The preparation of the stock solutions of the enzyme, the determinations of enzymatic activity and enzyme concentration, the methods of dissociation and reassociation, and the approaches to analyze the kinetics of reactivation and renaturation are also described in the preceding paper. Znz+ determination made use of the atomic absorption at 213.86 nm [ 2 ] . To provide comparable experimental conditions, 0.5 mg/ml bovine serum albumin or ribonuclease were added to the standard solutions with defined Zn2+ content.…”

Section: Methodsmentioning

confidence: 99%

Effect of Zinc(II) on the Refolding and Reactivation of Liver Alcohol Dehydrogenase

Rudolph

Gerschitz

Jaenicke

1978

European Journal of Biochemistry

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Horse-liver alcohol dehydrogenase requires Zn2' for enzymatic activity. Reactivation experiments after dissociation and denaturation of the enzyme in 6 M guanidinium hydrochloride and subsequent separation of zinc prove that the effect of the metal on the rate and yield of reconstitution is complex. In the absence of Zn2+ no reactivation is detectable, while excess of Zn2' leads to inactive aggregates. Optimum reactivation yields are obtained at 10 pM Zn2+ after short incubation in the denaturant; increasing zinc concentration causes a decrease of the rate of reactivation.The refolding of the zinc-free enzyme is characterized by consecutive first-order processes which may be separated from second-order dimer formation. Addition of 10 pM Zn2' during refolding may be used to block side reactions competing with the reconstitution. The transition from sigmoidal kinetics to second-order profiles by adding Zn2 * after completion of the aforementioned first-order process corroborates the proposed uni-bimolecular reactivation mechanism which implies the involvement of inactive monomers. These gain their enzymatic function as a consequence of dimerization.The effect of Zn2 ' may be explained by a side reaction in the overall reaction scheme of reactivation and renaturation which allows the kinetic measurements to be quantitatively described.In the preceding paper [l] equilibrium and kinetic studies on the reconstitution of horse-liver alcohol dehydrogenase after dissociation and denaturation in guanidinium hydrochloride were reported. Systematic variation of solvent parameters led to a high yield of reactivation and to a sufficient stability of the enzyme to allow a qualitative kinetic analysis of reactivation and renaturatjon. As shown by concentration-dependent experiments under essentially irreversible conditions, the sigmoidal traces of reactivation suggest a superposition of first-order transconformation and second-order association reactions. The coenzyme, NAD', does not show a significant influence on the kinetics of reactivation. Contrary to this, the presence of Zn2' (which is essential for the native structure, as well as the catalytic function of the enzyme) is shown to be important. Increasing concentrations of Zn2+ beyond the stoichiometric amount of two Zn2+ ions per monomer lead to a significant decrease in the Dedicated to Professor Theodor Wieland on the occasion of his 65th birthday.

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Section: Methodsmentioning

confidence: 99%

Effect of Zinc(II) on the Refolding and Reactivation of Liver Alcohol Dehydrogenase

Rudolph

Gerschitz

Jaenicke

1978

European Journal of Biochemistry

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“…Protective effect of various sera on cell damage induced by HX -X O Under conditions as described in the previous experiment, PCS and sera of normal (6), pregnant (6), and inflammatory (6) origins were compared for the protective effect against HX-X0.…”

Section: Resultsmentioning

confidence: 99%

“…(ICSH 1971) ; copper (Cu), direct bathocuproin method (Cu direct kit, Sinotest Lab.) (Matsubara 1967) ; zinc (Zn), atomic absorption spectrophotometric method using a Perkin Elmer atomic absorption spectrophotometer 370 (Fuwa et al 1968). Serum Cp, Tf, Hp, and crl acid glycoprotein (arl AG) were measured by single radial immunodifusion (M-Partigen kit, Hoechst) (Mancini 1965), and serum Hb was measured by SLS hemoglobin method (Hemoglobin B Test, Wako) (Ooki et al 1979).…”

Section: Measurement Of Serum Metals and Proteinsmentioning

confidence: 99%

Protective effect of the serum against cellular damage by active oxygen in culture.

Ogasawara

Kan

1984

Tohoku J. Exp. Med.

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“…Phosphate was measured by the method of Ames and Dubin (1). Atomic absorption was used for zinc measurements (7,8); standards were prepared from a Specpure zinc rod (Jarrell-Ash Co.) dissolved in dilute metal-free HCl and diluted with deionized water.…”

Section: Methodsmentioning

confidence: 99%

A Role for Zinc in the Structural Integrity of the Cytoplasmic Ribosomes of Euglena gacilis

Prask

Plocke

1971

Plant Physiol.

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Zinc deficiency in dark-grown Euglena gracilis Klebs, Z strain Pringshein, results in the disappearance of cytoplasmic ribosomes. In contrast, ribosomes in zinc-sufficient Euglena are conserved, do not undergo turnover, and can be demonstrated at any stage of growth. The zinc content of ribosomes from zincdeficient Euglena just prior to ribosomal disappearance is 300 to 380 micrograms of zinc per gram rRNA as compared to 650 to 1280 micrograms of zinc per gram rRNA in ribosomes from zinc-sufficient cells. Ribosomal disappearance is believed to involve a generalized disintegration process related to the lower content of zinc in the ribosomes. Reappearance of ribosomes requires the addition of zinc. It is proposed that adequate zinc may be essential for nornal tertiary and quaternary structure of the cytoplasmic ribosomes of Euglena.

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Determination of Zinc in Biological Materials by Atomic Absorption Spectrometry.

Cited by 148 publications

References 19 publications

Effect of Zinc(II) on the Refolding and Reactivation of Liver Alcohol Dehydrogenase

Effect of Zinc(II) on the Refolding and Reactivation of Liver Alcohol Dehydrogenase

Protective effect of the serum against cellular damage by active oxygen in culture.

A Role for Zinc in the Structural Integrity of the Cytoplasmic Ribosomes of Euglena gacilis

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