Ruthenium(II) tris(bathophenanthroline disulfonate)
Encyclopedia
Tetrasodium trisruthenium(II) is a coordination compound containing a ruthenium
center. In this form, it is the salt of a sulfonic acid
. This compound is an extension of the well known phenanthroline
series of coordination compounds. Ruthenium(II) tris(bathophenanthroline disulfonate), referring to the anionic fragment, is used as a protein
dye
in biochemistry
for differentiating and detecting different proteins in laboratory settings.
In recent years, 2-D electrophoresis has been widely accepted as a standard procedure to separate
complex protein mixtures in proteome studies (Proteomics
). Protein visualisation by Ruthenium(II) tris(bathophenthroline disulfonate) has become a firmly established and widely used method in proteomic analysis and a crucial step in protein expression
profiling.
For protein detection
, it is advantageous to use fluorescent labels containing chromophore
s which have longer excitation wavelength and emission wavelength than the aromatic amino acid
s. The dyes used for this important step should combine attributes like good signal to background ratio
(contrast), broad linear dynamic range, broad application range, photochemical stability and compatibility to protein identification techniques, e.g. mass spectrometry
(MS) or Western blotting.
Originally, the ruthenium
transition metal complex, ruthenium(II) tris(4,7-diphenyl-1,10-phenanthroline disulfonate) also termed as ruthenium(II) tris(bathophentroline disulfonate
) (RuBPS) was synthesized by Bannwarth http://www.chemie.uni-freiburg.de/orgbio/w3bann/index.html as a precursor molecule
for a dye that was used as a non-radioactive label for oligo
nucleotide
s. Later, Rabilloud et al. used RuBPS as a fluorescent label for protein detection in polyacrylamide
gel
s http://192.129.24.144/licensed_materials/10216/contents/00/00002/fpaper/s102160000002ch000.html. The fact that RuBPS is not only easy to synthesize but also easy to handle, induced further developments in this field.
Lamanda et al. improved the RuBPS staining protocol by selectively destaining the polyacrylamide matrix while the protein content remained tincture
d. This new technique entailed a variety of advantages like strong signals, ameliorated signal to background ratio, better linearity and advanced baseline resolution.
Ruthenium
Ruthenium is a chemical element with symbol Ru and atomic number 44. It is a rare transition metal belonging to the platinum group of the periodic table. Like the other metals of the platinum group, ruthenium is inert to most chemicals. The Russian scientist Karl Ernst Claus discovered the element...
center. In this form, it is the salt of a sulfonic acid
Sulfonic acid
Sulfonic acid usually refers to a member of the class of organosulfur compounds with the general formula RS2–OH, where R is an alkyl or aryl. The formal part of acid, HS2–OH, are formally derivatives of the "parent" inorganic compound with the formula HSO2.-Preparation:Sulfonic acid is...
. This compound is an extension of the well known phenanthroline
Phenanthroline
Phenanthroline is a heterocyclic organic compound. As a bidentate ligand in coordination chemistry, it forms strong complexes with most metal ions...
series of coordination compounds. Ruthenium(II) tris(bathophenanthroline disulfonate), referring to the anionic fragment, is used as a protein
Protein
Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
dye
Dye
A dye is a colored substance that has an affinity to the substrate to which it is being applied. The dye is generally applied in an aqueous solution, and requires a mordant to improve the fastness of the dye on the fiber....
in biochemistry
Biochemistry
Biochemistry, sometimes called biological chemistry, is the study of chemical processes in living organisms, including, but not limited to, living matter. Biochemistry governs all living organisms and living processes...
for differentiating and detecting different proteins in laboratory settings.
In recent years, 2-D electrophoresis has been widely accepted as a standard procedure to separate
Separation process
In chemistry and chemical engineering, a separation process, or simply a separation, is any mass transfer process used to convert a mixture of substances into two or more distinct product mixtures, at least one of which is enriched in one or more of the mixture's constituents. In some cases, a...
complex protein mixtures in proteome studies (Proteomics
Proteomics
Proteomics is the large-scale study of proteins, particularly their structures and functions. Proteins are vital parts of living organisms, as they are the main components of the physiological metabolic pathways of cells. The term "proteomics" was first coined in 1997 to make an analogy with...
). Protein visualisation by Ruthenium(II) tris(bathophenthroline disulfonate) has become a firmly established and widely used method in proteomic analysis and a crucial step in protein expression
Protein expression
Protein expression is a subcomponent of gene expression. It consists of the stages after DNA has been translated into polypeptide chains, which are ultimately folded into proteins...
profiling.
For protein detection
Spectroscopy
Spectroscopy is the study of the interaction between matter and radiated energy. Historically, spectroscopy originated through the study of visible light dispersed according to its wavelength, e.g., by a prism. Later the concept was expanded greatly to comprise any interaction with radiative...
, it is advantageous to use fluorescent labels containing chromophore
Chromophore
A chromophore is the part of a molecule responsible for its color. The color arises when a molecule absorbs certain wavelengths of visible light and transmits or reflects others. The chromophore is a region in the molecule where the energy difference between two different molecular orbitals falls...
s which have longer excitation wavelength and emission wavelength than the aromatic amino acid
Amino acid
Amino acids are molecules containing an amine group, a carboxylic acid group and a side-chain that varies between different amino acids. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen...
s. The dyes used for this important step should combine attributes like good signal to background ratio
Signal-to-noise ratio
Signal-to-noise ratio is a measure used in science and engineering that compares the level of a desired signal to the level of background noise. It is defined as the ratio of signal power to the noise power. A ratio higher than 1:1 indicates more signal than noise...
(contrast), broad linear dynamic range, broad application range, photochemical stability and compatibility to protein identification techniques, e.g. mass spectrometry
Mass spectrometry
Mass spectrometry is an analytical technique that measures the mass-to-charge ratio of charged particles.It is used for determining masses of particles, for determining the elemental composition of a sample or molecule, and for elucidating the chemical structures of molecules, such as peptides and...
(MS) or Western blotting.
Originally, the ruthenium
Ruthenium
Ruthenium is a chemical element with symbol Ru and atomic number 44. It is a rare transition metal belonging to the platinum group of the periodic table. Like the other metals of the platinum group, ruthenium is inert to most chemicals. The Russian scientist Karl Ernst Claus discovered the element...
transition metal complex, ruthenium(II) tris(4,7-diphenyl-1,10-phenanthroline disulfonate) also termed as ruthenium(II) tris(bathophentroline disulfonate
Sulfonate
A sulfonate is a salt or ester of a sulfonic acid. It contains the functional group R-SO2O-.- Sulfonate salts:Anions with the general formula RSO2O− are called sulfonates. They are the conjugate bases of sulfonic acids with formula RSO2OH. As sulfonic acids tend to be strong acids, the...
) (RuBPS) was synthesized by Bannwarth http://www.chemie.uni-freiburg.de/orgbio/w3bann/index.html as a precursor molecule
Precursor (chemistry)
In chemistry, a precursor is a compound that participates in the chemical reaction that produces another compound. In biochemistry, the term "precursor" is used more specifically to refer to a chemical compound preceding another in a metabolic pathway....
for a dye that was used as a non-radioactive label for oligo
Oligonucleotide
An oligonucleotide is a short nucleic acid polymer, typically with fifty or fewer bases. Although they can be formed by bond cleavage of longer segments, they are now more commonly synthesized, in a sequence-specific manner, from individual nucleoside phosphoramidites...
nucleotide
Nucleotide
Nucleotides are molecules that, when joined together, make up the structural units of RNA and DNA. In addition, nucleotides participate in cellular signaling , and are incorporated into important cofactors of enzymatic reactions...
s. Later, Rabilloud et al. used RuBPS as a fluorescent label for protein detection in polyacrylamide
Polyacrylamide
Polyacrylamide is a polymer formed from acrylamide subunits. It can be synthesized as a simple linear-chain structure or cross-linked, typically using N,N-methylenebisacrylamide. Polyacrylamide is not toxic...
gel
Gel
A gel is a solid, jelly-like material that can have properties ranging from soft and weak to hard and tough. Gels are defined as a substantially dilute cross-linked system, which exhibits no flow when in the steady-state...
s http://192.129.24.144/licensed_materials/10216/contents/00/00002/fpaper/s102160000002ch000.html. The fact that RuBPS is not only easy to synthesize but also easy to handle, induced further developments in this field.
Lamanda et al. improved the RuBPS staining protocol by selectively destaining the polyacrylamide matrix while the protein content remained tincture
Tincture
A tincture is an alcoholic extract or solution of a non-volatile substance . To qualify as a tincture, the alcoholic extract is to have an ethanol percentage of at least 40-60%...
d. This new technique entailed a variety of advantages like strong signals, ameliorated signal to background ratio, better linearity and advanced baseline resolution.