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Synthesis of 1,4‐disubstituted aminoanthraquinone derivatives for purification of lactate dehydrogenase
Author(s) -
Kubista Jordan Louise
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.784.4
Subject(s) - anthraquinone , lactate dehydrogenase , chemistry , moiety , combinatorial chemistry , quenching (fluorescence) , dehydrogenase , nad+ kinase , enzyme , stereochemistry , organic chemistry , fluorescence , physics , quantum mechanics
Cibacron Blue 3G‐A is a dye commonly used for the purification of various enzymes, in particular, isolating lactate dehydrogenase. Companies have Cibacron Blue available periodically but it is rather expensive. Because of these problems, an alternative dye that can be easily synthesized and has similar properties to Cibacron Blue is desired. Cibacron Blue is a dye designed to mimic natural biological ligands, such as NAD + /NADH. A suitable replacement should exhibit comparable affinity and purifying ability for targeted proteins. Cibacron Blue contains an anthraquinone core, which is a distinct feature due to its versatility. Many different functional groups can be substituted on to the anthraquinone core giving it a multitude of different properties and functions. The goal of this research is to synthesize an anthraquinone derivative that exhibits similar properties and functions as Cibacron Blue. The alternatives include aromatic anilines at the 1,4 positions of the anthraquinone to emulate Cibacron Blue's structure at the anthraquinone core and allow for attachment to a solid support. These were synthesized by treatment of 1,4‐bistosyloxyanthraquinone with two substituted anilino nucleophiles to build a library. These alternatives should mimic the adenine moiety of NAD + , which are accommodated in a hydrophobic crevice formed by five chain regions of lactate dehydrogenase. The left side of the anthraquinone is left unsubstituted since it is believed this is where it binds to the hydrophobic nucleic fold of enzymes. After the synthesis of the small library of aminoanthraquinone derivatives was completed, they are characterized. Binding affinity was tested using Fluorescence quenching. This method allows for comparisons between the binding of lactate dehydrogenase to the synthesized products and binding of lactate dehydrogenase to NADH using K d values. The K d value is an equilibrium dissociation constant that is used to evaluate binding affinity. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .