DNA-maleimide: an improved maleimide compound for electrophoresis-based titration of reactive thiols in a specific protein

Satoshi Hara; Tatsuya Nojima; Kohji Seio; Masasuke Yoshida; Toru Hisabori

doi:10.1016/j.bbagen.2013.01.012

DNA-maleimide: an improved maleimide compound for electrophoresis-based titration of reactive thiols in a specific protein

Biochim Biophys Acta. 2013 Apr;1830(4):3077-81. doi: 10.1016/j.bbagen.2013.01.012. Epub 2013 Jan 26.

Authors

Satoshi Hara¹, Tatsuya Nojima, Kohji Seio, Masasuke Yoshida, Toru Hisabori

Affiliation

¹ Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-ku, Yokohama 226-8503, Japan.

PMID: 23357041
DOI: 10.1016/j.bbagen.2013.01.012

Abstract

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.

General significance: The versatility of DNA molecule enhances the application of DNA-Mal in a broader range of cysteine containing proteins.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA, Single-Stranded / chemistry*
Electrophoresis / methods*
Maleimides / chemistry*
Sulfhydryl Compounds / analysis*
Sulfhydryl Reagents / chemistry*

Substances

DNA, Single-Stranded
Maleimides
Sulfhydryl Compounds
Sulfhydryl Reagents
maleimide