Disulfide Bond Analysis

Introduction
Disulfide bond is a single covalent bond formed between the sulfur atoms of cysteines. The
other sulfur-containing amino acid, methionine, cannot form disulfide bonds. Because it is a
covalent bond, disulfide bond is often considered to be the primary structure. However, the
function of disulfide bonds are far more than components of primary protein structure, they
play a very important role in stabilizing the tertiary and quartenary structures, and are the
prerequisite of proteins’ proper biological function.

Function of disulfide bonds

 At tertiary structure level: Disulfide bonds are formed inside the protein subunit at this level.
Disulfide bonds play an important role in the folding and stability of some proteins, usually
before proteins are secreted to the extracellular medium. The disulfide bond may form the
nucleus of a hydrophobic core of the folded protein, i.e., local hydrophobic residues may
condense around the disulfide bond and onto each other through hydrophobic interactions.
 At quartenary structure level: Disulfide bonds are formed between two protein subunits. This
type of disulfide bonds hold two portions of the protein together, biasing the protein towards
the folded topology. That is, the disulfide bond destabilizes the unfolded form of the protein by
lowering its entropy. The disulfide bond increases the effective local concentration of protein
residues and lowers the effective local concentration of water molecules. Since water
molecules attack amide-amide hydrogen bonds and break up secondary structure, a disulfide
bond stabilizes secondary structure in its vicinity. For example, researchers have identified
several pairs of peptides that are unstructured in isolation, but adopt stable secondary and
tertiary structure upon forming a disulfide bond between them.
Given the ability of stabilizing overall structure of proteins, disulfide bridges are cross-linked in
many commercialized proteins to increase their resistance to destructive effects of extreme
environment used in industrial processes or protect protein-based therapeutics from rapid
proteolytic degradation. Manufacturing of these products must take into account oxidative
refolding—a formation of native disulfide bonds by specific pairs of cysteines located
throughout a sequence of linear protein.
 Creative Proteomics has established a highly sensitive HPLC-MS/MS platform that can
analyze disulfide bonds in multiple samples and in both eukaryotic and prokaryotic organisms.
In addition, we have optimized our protocol to enable more fast and sensitive site mapping
service for disulfide bond analysis.

Applications of disulfide bond analysis service:

 Study unknown disulfide bonds in novel proteins.
 Analyze disulfide bonds in refolded proteins to test whether a protein is correctly folded.
Technology platform:
 Ion Chromatography
 High Performance Liquid Chromatography (HPLC)
 Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS)
Creative Proteomics also provide the following bioinformatics services in Protein Post-
translational Modification Analysis:
Functional annotation and enrichment analysis
Clustering analysis
Network analysis
Statistical analysis
Proteomic analysis of post-translational modifications
Please feel free to Contact Us to discuss your projects. We hope you will find that we can
meet your research needs.