Witters Erwin, Quanten Lieve, Bloemen Jo, Valcke Roland, Van Onckelen Harry
Laboratory for Plant Biochemistry and Plant Physiology, Department of Biology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
Rapid Commun Mass Spectrom. 2004;18(4):499-504. doi: 10.1002/rcm.1365.
In view of the ongoing debate on plant cyclic nucleotide metabolism, especially the functional presence of adenylyl cyclase, a novel detection method has been worked out to quantify the reaction product. Using uniformly labelled (15)N-ATP as a substrate for adenylyl cyclase, a qualitative and quantitative liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) method was developed to measure de novo formed (15)N-adenosine 3',5'-cyclic monophosphate. Adenylyl cyclase activity was observed in chloroplasts obtained from Nicotiana tabacum cv. Petit Havana and the kinetic parameters and influence of various metabolic effectors are discussed in their context.
鉴于目前关于植物环核苷酸代谢的争论，尤其是腺苷酸环化酶的功能存在情况，已开发出一种新的检测方法来定量反应产物。使用均匀标记的(15)N-ATP作为腺苷酸环化酶的底物，开发了一种定性和定量的液相色谱/电喷雾电离串联质谱(LC/ESI-MS/MS)方法来测量新形成的(15)N-腺苷3',5'-环一磷酸。在烟草品种 Petit Havana 的叶绿体中观察到了腺苷酸环化酶活性，并在相关背景下讨论了动力学参数和各种代谢效应物的影响。
