Materials Chemistry and Chemical Analysis, University of Turku, Vatselankatu 2, 20500 Turku, Finland.
NCI-RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD 21702, USA.
Sensors (Basel). 2018 Nov 16;18(11):3989. doi: 10.3390/s18113989.
A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in their hydrolysis assays performed in a high throughput screening compatible 384-well plate format.
本文提出了一种用于在中性水溶液中检测发光核苷酸的新型无标记分子探针。含磷分子，如核苷酸，在细胞代谢、能量经济和各种信号过程中起着至关重要的作用。因此，监测核苷酸浓度和与核苷酸相关的酶反应非常重要。由 Tb(III)离子载体和光收集天线组成的两部分镧系元素配合物，很容易区分含有不同数量磷酸的核苷酸，并能够直接检测将三磷酸核苷酸（NTP）转化为二/单磷酸核苷酸的酶反应，反之亦然。开发的传感器能够以低纳摩尔的灵敏度检测酶活性，在以高通量筛选兼容的 384 孔板格式进行的 K-Ras 和脱氨酶酶水解测定中得到了突出体现。
