AN IN VITRO MICRO-VOLUME PROCEDURE FOR RAPID MEASUREMENT OF ERYTHROCYTIC HEXOSE MONOPHOSPHATE SHUNT ACTIVITY

Authors

J. Kevin Baird, ALERTAsia FoundationFollow
Joan E. Decker-Jackson, Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Walter Reed Army Medical Center, Washington, DC
David E. Davidson Jr., Division of Experimental Therapeutics, Walter Reed Army Institute of Research, Walter Reed Army Medical Center, Washington, DC

Date of this Version

1984

Citation

Inl. J. Biochem, Vol. 16, No. 10, pp. 1049-1052, 1984

Comments

U.S. Government Work

Abstract

I. A radiometric micro-volume procedure for measurement of erythrocytic hexose monophosphate shunt (HMS) activity in intact cells in vitro is described.

2.The procedure is rapid, allowing 200 individual HMS determinations in a single experiment of 5 hr duration.

3. The procedure is reproducible, yielding HMS activity means insignificantly different (P > 0.05) between replicate experiments.

4. A profile of sodium nitrite-induced HMS stimulation is reported: HMS was elevated 2-fold (P < 0.001) between zero and 2.5mM NaN02; HMS elevation was more distinct (7-fold) between 2.5 and 5.0mM NaNO₂; maximum activity (22-fold) was observed between 10 and 20mM NaN02; > 20mM NaNO₂ caused significant (P < 0.001) diminution of HMS; glucose carbon recycling through the HMS occurred only with > 2.5mM NaNO₂ where this process contributed :::; 16% to total HMS activity.