N-hydroxysulfosuccinimide active esters: bis(N-hydroxysulfosuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane-impermeant, protein cross-linkers

Biochemistry. 1982 Aug 17;21(17):3950-5. doi: 10.1021/bi00260a008.


We have synthesized and characterized N-hydroxysulfosuccinimide, a new hydrophilic ligand for the preparation of active esters. We have incorporated this ligand into two new protein cross-linking reagents, 3,3'-dithiobis(sulfosuccinimidyl propionate) and bis(sulfosuccinimidyl) suberate. In experiments with rabbit muscle aldolase, it is demonstrated that both of these reagents are highly efficient protein cross-linkers at physiological pH and that 3,3'-dithiobis(sulfosuccinimidyl propionate) is quantitatively cleavable by reduction under mild conditions. In experiments with intact human erythrocytes and erythrocyte membranes, it is shown that both reagents are membrane impermeant and that when erythrocytes are treated with either reagent, both cross-link subunits of the anion channel (band 3) to covalent dimers at the extracytoplasmic membrane face.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Membrane Permeability*
  • Chemical Phenomena
  • Chemistry
  • Cross-Linking Reagents* / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Erythrocyte Membrane / drug effects
  • Erythrocytes / drug effects
  • Fructose-Bisphosphate Aldolase
  • Macromolecular Substances
  • Muscles / enzymology
  • Succinimides*


  • Cross-Linking Reagents
  • Macromolecular Substances
  • Succinimides
  • 3,3'-dithiobis(sulfosuccinimidyl propionate)
  • N-hydroxysulfosuccinimide
  • bis(sulfosuccinimidyl)suberate
  • Fructose-Bisphosphate Aldolase