Reagents for rapid reduction of disulfide bonds Book

Singh, R, Lamoureux, GV, Lees, WJ et al. (1995). Reagents for rapid reduction of disulfide bonds . 251 167-173. 10.1016/0076-6879(95)51119-9



cited authors

  • Singh, R; Lamoureux, GV; Lees, WJ; Whitesides, GM

fiu authors

abstract

  • This chapter describes the reagents that reduce disulfide bonds, under nondenaturing conditions at pH 7, faster than does dithiothreitol (DTT) by a factor of ∼5–7. Dithiothreitol and 2-mercaptoethanol (ME) are the most commonly used reagents for the reduction of disulfide bonds. However, both DTT and ME have drawbacks in their use: DTT is slow in reducing disulfide bonds at pH 7; ME is foul smelling and may not completely reduce disulfide bonds owing to its low reduction potential. A new class of dithiol reagents for rapid reduction of disulfide bonds at pH 7 to 8 is based on two requirements: (1) a low pKa value (∼7–8) for their thiol groups, so that a significant proportion of the thiol is in the reactive thiolate form and (2) a high value for their reduction potential. These dithiol reagents reduce disulfide bonds by the mechanism of thiol–disulfide interchange. The chapter describes three reagents—bis(2-mercaptoethyl) sulfone (BMS), meso-2,5-dimercapto-N,N,N',N'-tetramethyladipamide (DTA), and N,N'-dimethyl-N,N'-bis(mercaptoacetyl)hydrazine (DMH)—that have the desirable properties of high reduction potential and pKa values of ∼7.8. These dithiol reagents (BMS, DTA, and DMH) reduce disulfide bonds in proteins under nondenaturing conditions at pH 7 faster than dithiothreitol. All three reagents are conveniently synthesized from readily available materials. © 1995 Elsevier Inc.

publication date

  • January 1, 1995

Digital Object Identifier (DOI)

start page

  • 167

end page

  • 173

volume

  • 251