PMID: 45777Jan 1, 1978

Binding of mercurials to membrane suspensions and undenatured proteins

Membrane Biochemistry
G G Berg, E F Miles

Abstract

Binding capacities of membrane suspensions and dissolved compounds for mercurials were titrated by a new potentiometric method. Critical steps included a silver electrode of new design, the use of L-cysteine as a thiol buffer, a nitrogen atmosphere, and pretreatment of samples with equimolar mercurial and cysteine. Titrations had a sharp endpoint, accurate +/- 26 nmole methylmercury or +/- 8 nmole mercuric salt. Measurements of binding capacity of bovine serum albumin averaged 93% of the titer predicted for one SH group per molecule; those of human hemoglobin yielded 86-91% of the titer predicted for two SH groups per molecule. Yields dropped with exposure of protein solutions or membrane suspensions to atmospheric oxygen. Brain microsomes had significantly higher binding capacities (per milligram of protein) than red blood cell ghosts. The ratio of endpoint titers of CH3HgCl to HgCl2 averaged 2:1 in assays of cysteine, proteins, and membranes, showing that the assay was free of denaturation artifacts and protein-protein interference. Solutions of EDTA showed measurable binding of Hg2+ but not of CH3Hg+. Satisfactory titrations were also obtained with N-ethylmaleimide.

References

Jan 1, 1975·Journal of Supramolecular Structure·F H KirkpatrickR I WEED
Jan 15, 1973·European Journal of Biochemistry·M H FreedmanJ S Cohen
Feb 1, 1974·Analytical Chemistry·P W Alexander, G A Rechnitz
Jun 26, 1972·Biochimica Et Biophysica Acta·J M Rifkind
Dec 1, 1968·Biochemistry·P G SquireC T O'Konski
Apr 11, 1957·Annals of the New York Academy of Sciences·W L HUGHES
May 1, 1958·The Biochemical Journal·A C ALLISON, R CECIL
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN
Jan 1, 1962·The Journal of General Physiology·R I WEEDA ROTHSTEIN
Mar 1, 1965·The Journal of General Physiology·J VANSTEVENINCKA ROTHSTEIN
Oct 1, 1970·Talanta·T Y Tombara, L Koval
Oct 1, 1970·Talanta·T Y ToribaraL Koval

Citations

Oct 1, 1979·Chemico-biological Interactions·G G Berg, E F Miles
Sep 23, 1997·Toxicology and Applied Pharmacology·G G Berg

Related Concepts

Brain
Plasma Membrane
Electrode
Red Cell Ghost
Microsomes
Potentiometry
Protein Denaturation
Serum Albumin, Bovine
Sulfhydryl Compounds
Mercury Compounds, Inorganic

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