NDLI: Calculation of the Effect of Solanine on the Michaelis Constant and the Maximum Reaction Rate of NAT

Content Provider	IEEE Xplore Digital Library
Author	Gao Shi-yong Ji Yu-bin Zou Xiang Ji Chen-feng
Copyright Year	2008
Description	Author affiliation: Center of R&D on Life Sci. & Environ. Sci., Harbin Univ. of Commerce, Harbin (Gao Shi-yong; Ji Yu-bin; Zou Xiang; Ji Chen-feng)
Abstract	To explore how to calculate the effect of solanine on the Michaelis constant and the maximum reaction rate of NAT, high performance liquid chromatography (HPLC) was used, with 2-AF as substrate, and the rate at which 2-AF is acetylated into 2-AAF in intact $HepG_{2}$ cells or in the cytoplasm of $HepG_{2}$ cells as the reaction rate. The double reciprocal plot was made, with 1/S (reciprocal of the concentration of the substrate 2-AF) plotted against 1/V (reciprocal of the reaction rate), to yield a regression equation for calculating $K_{m}$ and $V_{max}.$ With intact $HepG_{2}$ cells, the $K_{m}$ and $V_{max}$ for the negative control are $2.37times10^{-3}plusmn8.37times10^{-5}$ mM and $9.16times10^{-4}plusmn7.54times10^{-5}$ $nmol/10^{6}$ cells, respectively, and that the $K_{m}$ and $V_{max}$ for the solanine group are $2.22times10^{-3}plusmn9.05times10^{-5}$ mM and $5.14times10^{-4}plusmn3.72times10^{-5}$ $nmol/10^{6}$ cells, respectively. For the cytoplasm of $HepG_{2}$ cells, the $K_{m}$ and $V_{max}$ for the negative control are $8.95times10^{-3}plusmn2.61times10^{-4}$ mM and $2.55times10^{-6}plusmn1.92times10^{-8}$ nmol/ minmiddotmg protein, respectively, and the $K_{m}$ and $V_{max}$ of the solanine group are $9.48times10^{-3}plusmn3.63times10^{-4}$ Mm and $2.43times10^{-6}plusmn1.32times10^{-8}$ nmol/minmiddotmg protein. Statistical analysis showed that, for both intact $HepG_{2}$ cells and cytoplasm of $HepG_{2}$ cells, $K_{m}$ does not differ significantly between the negative control and the solanine groups, but $V_{max}$ does differ significantly for these groups (P<0.05). Solanine is a non-competitive inhibitor for the 2-AF substrate of NAT.
Starting Page	1260
Ending Page	1263
File Size	168421
Page Count	4
File Format	PDF
ISBN	9781424417476
DOI	10.1109/ICBBE.2008.643
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2008-05-16
Publisher Place	China
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Proteins Inhibitors Chemical analysis Bovine Kinetic theory Pancreas Network address translation Neoplasms Research and development Cancer
Content Type	Text
Resource Type	Article

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