Simultaneous measurement of CYP1A2 activity, regioselectivity, and coupling: Implications for environmental sensitivity of enzyme-substrate binding.
Simultaneous measurement of CYP1A2 activity, regioselectivity, and coupling: Implications for environmental sensitivity of enzyme-substrate binding.
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同时测量 CYP1A2 活性、区域选择性和偶联：对酶-底物结合的环境敏感性的影响。
DOI：
10.1016/j.abb.2010.10.002
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发表时间：
2011
影响因子：
3.9
通讯作者：
Clark,DouglasS
中科院分区：
文献类型：
--
作者：
Traylor,MatthewJ;Chai,Jack;Clark,DouglasS
The cytochrome P450 (CYP) reaction mechanism often yields a broad array of coupled and uncoupled products from a single substrate. While it is well known that reaction conditions can drastically affect the rate of P450 catalysis, their effects on regioselectivity and coupling are not well characterized. To investigate such effects, the CYP1A2 oxidation of 7-ethoxymethoxy-3-cyanocoumarin (EOMCC) was examined as a function of buffer type, buffer concentration, pH, and temperature. A high-throughput, optical method was developed to simultaneously measure the rate of substrate depletion, NADPH depletion, and generation of the O-dealkylated product. Increasing the phosphate buffer concentration and temperature increased both the NADPH and EOMCC depletion rates by 6-fold, whereas coupling was constant at 7.9% and the regioselectivity of O-dealkylation to other coupled pathways was constant at 21.7%. Varying the buffer type and pH increased NADPH depletion by 2.5-fold and EOMCC depletion by 3.5-fold; however, neither coupling nor regioselectivity was constant, with variations of 14.4% and 21.6%, respectively. Because the enzyme–substrate binding interaction is a primary determinant of both coupling and regioselectivity, it is reasonable to conclude that ionic strength, as varied by the buffer concentration, and temperature alter the rate without affecting binding while buffer type and pH alter both.
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影响因子：
3.9
作者：
H. Yamazaki;E. Gillam;M. Dong;William W. Johnson;F. Guengerich;T. Shimada
通讯作者：
H. Yamazaki;E. Gillam;M. Dong;William W. Johnson;F. Guengerich;T. Shimada
碳酸根阴离子对细胞色素 P450 2D6 介导的体外代谢的影响：多种氧化物质的潜在作用。
影响因子：
3.9
作者：
J. Hutzler;Frank J Powers;M. Wynalda;L. Wienkers
通讯作者：
L. Wienkers
影响因子：
2.9
作者：
Makris, Thomas M.;von Koenig, Konstanze;Sligar, Stephen G.
通讯作者：
Sligar, Stephen G.
影响因子：
2.9
作者：
Kim, JS;Ahn, T;Yun, CH
通讯作者：
Yun, CH
人类细胞色素 P450 1A2 和某些突变体氧化苯乙酸的速率决定步骤
影响因子：
2.9
作者：
Yun, CH;Miller, GP;Guengerich, FP
通讯作者：
Guengerich, FP