Disuccinimidyl suberate (DSS) is a six-carbon
lysine
Lysine (symbol Lys or K) is an α-amino acid that is a precursor to many proteins. It contains an α-amino group (which is in the protonated form under biological conditions), an α-carboxylic acid group (which is in the deprotonated −C ...
-reactive non-cleavable
cross-linking agent.
It consists of functional groups It is a homobifunctional
N-hydroxysuccinimide (NHS) ester formed by carbodiimide-activation of carboxylate molecules, with identical reactive groups at either end. The reactive groups are separated by a spacer and in this molecule it is a six carbon alkyl chain.
This reagent is mainly used to form intramolecular crosslinks and preparation of
polymers
A polymer (; Greek '' poly-'', "many" + ''-mer'', "part")
is a substance or material consisting of very large molecules called macromolecules, composed of many repeating subunits. Due to their broad spectrum of properties, both synthetic an ...
from
monomers. It is ideal for receptor ligand cross-linking.
DSS is reactive towards
amine
In chemistry, amines (, ) are compounds and functional groups that contain a basic nitrogen atom with a lone pair. Amines are formally derivatives of ammonia (), wherein one or more hydrogen atoms have been replaced by a substituen ...
groups (primary amines) at pH 7.0-9.0. It is membrane permeable, therefore permitting intracellular cross-linking, has high purity, is non-cleavable, and is water-insoluble (it must be dissolved in a polar organic solvent such as DMF or DMSO before addition to sample.)
Its reaction specificity, reaction product stability, and lack of reaction by-products make it a commonly used cross-linking agent.
See also
*
Bissulfosuccinimidyl suberate, a water-soluble version
Applications
* Chemical crosslinking of intracellular proteins prior to cell lysis and immunoprecipitation
* 'Fix' protein interactions to allow identification of weak or transient protein interactions
* Protein crosslinking to create bioconjugates via single-step reactions
* Immobilize proteins onto amine-coated surfaces
* Crosslinking mass spectrometry (crosslinking-MS) provides insight into protein structure, organization, and interactions
Size Exclusion Chromatography
A study was done looking at the application of
size exclusion chromatography in the purification of cross-link peptides within samples. When the eluted peptides were analyzed, cross-linked peptides could be detected at higher concentrations in the earlier fractions, eluting before modified or unmodified peptides.
DSS Cross-linking with Hemoglobin-albumin
Disuccinimidyl suberate's reactivity toward primary amines allows it to serve as a cross-linking agent for proteins, without toxic side-products and forming peptide bonds with the lysine residues in a single step. In a study on blood substitutes, DSS was shown to cross-link
Hemoglobin
Hemoglobin (haemoglobin BrE) (from the Greek word αἷμα, ''haîma'' 'blood' + Latin ''globus'' 'ball, sphere' + ''-in'') (), abbreviated Hb or Hgb, is the iron-containing oxygen-transport metalloprotein present in red blood cells (erythrocyte ...
intramolecularly, yielding a relatively stable protein (polymerized Hb or polyHb), whose oxygen affinity was almost halved compared to that of native Hb. This was shown to be reversed when Hemoglobin was copolymerized with
bovine serum albumin
Bovine serum albumin (BSA or "Fraction V") is a serum albumin protein derived from cows. It is often used as a protein concentration standard in lab experiments.
The nickname "Fraction V" refers to albumin being the fifth fraction of the origin ...
(BSA), showing very little change in auto-oxidation and oxygen affinity compared to the native Hb.
References
Succinimides
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