DNA-maleimide: an improved maleimide compound for electrophoresis-based titration of reactive thiols in a specific protein.
BACKGROUND
Thiol-mediated redox regulation of proteins plays a key role in many cellular processes.


METHODS
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).


RESULTS
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.


CONCLUSIONS
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.


CONCLUSIONS
The versatility of DNA molecule enhances the application of DNA-Mal in a broader range of cysteine containing proteins.
Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-ku, Yokohama 226-8503, Japan.
Satoshi
S
Hara
Tatsuya
T
Nojima
Kohji
K
Seio
Masasuke
M
Yoshida
Toru
T
Hisabori
D004277:DNA, Single-Stranded / Q000737:chemistry*; D004586:Electrophoresis / Q000379:methods*; D008301:Maleimides / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000032:analysis*; D013439:Sulfhydryl Reagents / Q000737:chemistry*
D004277:DNA, Single-Stranded; D008301:Maleimides; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; C043592:maleimide