Probe compound for detecting and isolating enzymes and means and methods using the same
申请人:Helmholtz-Zentrum für Infektionsforschung GmbH
公开号:EP2230312A1
公开(公告)日:2010-09-22
The present invention relates to a probe compound that can comprise any substrate or metabolite of an enzymatic reaction in addition to an indicator component, such as, for example, a fluorescence dye, or the like. Moreover, the present invention relates to means for detecting enzymes in form of an array, which comprises any number of probe compounds of the invention which each comprise a different metabolite of interconnected metabolites representing the central pathways in all forms of life. Moreover, the present invention relates to a method for detecting enzymes involving the application of cell extracts or the like to the array of the invention which leads to reproducible enzymatic reactions with the substrates. These specific enzymatic reactions trigger the indicator (e.g. a fluorescence signal) and bind the enzymes to the respective cognate substrates. Moreover, the invention relates to means for isolating enzymes in form of nanoparticles coated with the probe compound of the invention. The immobilisation of the cognate substrates or metabolites on the surface of nanoparticles by means of the probe compounds allows capturing and isolating the respective enzyme, e.g. for subsequent sequencing.
本发明涉及一种探针化合物,它可以包括酶反应的任何底物或代谢物,此外还包括指示成分,例如荧光染料或类似物。此外,本发明还涉及以阵列形式检测酶的方法,该阵列由任意数量的本发明探针化合物组成,每种探针化合物由代表所有生命形式中中心途径的相互关联的代谢物中的不同代谢物组成。此外,本发明还涉及一种检测酶的方法,该方法涉及将细胞提取物或类似物应用于本发明的阵列,从而导致与底物发生可重复的酶反应。这些特定的酶反应会触发指示剂(如荧光信号),并将酶与各自的同源底物结合。此外,本发明还涉及以涂覆有本发明探针化合物的纳米颗粒形式分离酶的方法。通过探针化合物将同源底物或代谢物固定在纳米颗粒表面,可以捕获和分离相应的酶,例如用于后续测序。