Clinical Chemistry
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Clinical Chemistry 27: 1401-1409, 1981;
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Clinical Chemistry, Vol 27, 1401-1409, Copyright © 1981 by American Association for Clinical Chemistry

Measurement of alkaline phosphatase activity: characterization and identification of an inactivator in 2-amino-2-methyl-1-propanol

R Rej, JP Bretaudiere, RW Jenny and KY Jackson

An inactivator of alkaline phosphatase (EC 3.1.3.1) in 2-amino-2-methyl- 1-propanol is demonstrated and characterized. This time-dependent inactivation results from chelation of enzyme-bound Zn2+; it is reversed by addition of Zn2+ and, to a lesser extent, other divalent metal ions. Cu2+ is an effective spectral indicator and can be used to determine the presence and quantity of inactivator. Data obtained from enzyme inactivation, Cu2+ absorbance spectra, "high-performance" liquid chromatography, thin-layer chromatography, Fourier-transform infrared spectroscopy, and mass spectroscopy indicate that the inactivator is 5- amino-3-aza-2,2,5-trimethylhexanol. This compound, even in trace amounts (less than 0.05% on a molar basis), shown to inactivate alkaline phosphatase.




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Copyright © 1981 by the American Association for Clinical Chemistry.