Hara Satoshi, Nojima Tatsuya, Seio Kohji, Yoshida Masasuke, Hisabori Toru
Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-ku, Yokohama 226-8503, Japan.
Biochim Biophys Acta. 2013 Apr;1830(4):3077-81. doi: 10.1016/j.bbagen.2013.01.012. Epub 2013 Jan 26.
Thiol-mediated redox regulation of proteins plays a key role in many cellular processes.
To understand the redox status of cysteinyl thiol groups of the desired proteins, we developed a new maleimide reagent: a maleimide-conjugated single strand DNA, DNA-maleimide (DNA-Mal).
DNA-Mal labelled proteins run as a distinct band on SDS-PAGE, with a discrete 9.32 kDa mobility shift per label regardless of the protein species or electrophoretic conditions.
DNA-Mal labels free thiols like standard maleimide reagents, but possesses practical advantages in titration of the number and relative content of free thiols in a protein.
The versatility of DNA molecule enhances the application of DNA-Mal in a broader range of cysteine containing proteins.
蛋白质的硫醇介导的氧化还原调节在许多细胞过程中起关键作用。
为了解所需蛋白质的半胱氨酰硫醇基团的氧化还原状态，我们开发了一种新的马来酰亚胺试剂：马来酰亚胺共轭单链DNA，即DNA-马来酰亚胺（DNA-Mal）。
DNA-Mal标记的蛋白质在SDS-PAGE上呈现为一条独特的条带，无论蛋白质种类或电泳条件如何，每个标记的迁移率离散变化9.32 kDa。
DNA-Mal与标准马来酰亚胺试剂一样标记游离硫醇，但在滴定蛋白质中游离硫醇的数量和相对含量方面具有实际优势。
DNA分子的多功能性增强了DNA-Mal在更广泛的含半胱氨酸蛋白质中的应用。
