Biotin Hydrazide

Biotin hydrazide is a biotinyl derivative that can be used as a probe for the determination of protein carbonylation, which is a component of several diseases. Protein carbonylation, an irreversible post translational modification (PTM), is caused by attack of reactive oxygen species (ROS), numerous lipid oxidation products (such as α,β-unsaturated γ-hydroxyalkenals), or nonemzymatic glycation resulting in the loss of protein function. Biotin hydrazide is a preferred carbonyl-reactive probe for its direct reaction and chemistry and no requirement of additional catalysts or reducing agents. Biotin hydrazide exclusively and readily derivatizes carbonyl groups at pH 5.5, which facilitates its use to measure carbonylated proteins in biological samples.

Reference

Kenneth Hensley, Kelly S. Williamson. Protein Carbonyl Determination Using Biotin Hydrazide. Methods in Biological Oxidative Stress. Methods in Pharmacology and Toxicology 2003, pp 195-199

Madian AG, Regnier FE. Proteomic identification of carbonylated proteins and their oxidation sites. J Proteome Res. 2010;9(8):3766-3780.

Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide.[Pubmed:26590475]

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Apr 15;1019:128-31.

Protein carbonyls are protein oxidation products that are often used to measure the magnitude of protein oxidative damage induced by reactive oxygen or reactive nitrogen species. Protein carbonyls have been found to be elevated during aging and in age-related diseases such as stroke, diabetes, and neurodegenerative diseases. In the present article, we provide detailed protocols for detection of mitochondrial protein carbonyls labeled with biotin-hydrazide followed by 2-dimensional isoelectric focusing (IEF)/SDS-PAGE and Western blotting probed with horse-radish peroxidase-conjugated streptavidin. The presented procedures can also be modified for detection of carbonylation of non-mitochondrial proteins.

Comparative electrochemical behaviour of biotin hydrazide and photobiotin. Importance in the development of biosensors.[Pubmed:10641292]

Biosens Bioelectron. 1999 Dec;14(8-9):729-35.

The cyclic voltammetric behaviour of Biotin Hydrazide and photobiotin on carbon paste electrodes has been studied. Biotin Hydrazide presents an anodic and irreversible process, meanwhile photobiotin presents two, adsorptive in nature. This characteristic makes photobiotin desirable for following the interaction between biotin and streptavidin, being possible to detect a streptavidin concentration of 10(-12) M. The evidence of this reaction has been shown either directly in solution or on the electrode surface. Photobiotin as the molecule portable of analytical information and carbon paste as the solid support could be applied to the development of sensors based on the oxidation of this molecule.

Detection of Protein Carbonyls by Means of Biotin Hydrazide-Streptavidin Affinity Methods.[Pubmed:26139258]

Methods Mol Biol. 2015;1314:95-100.

Oxidative posttranslational protein modifications occur as a normal process of cell biology and to a greater extent during pathogenic conditions. The detection and quantitation of protein oxidation has posed a continuing challenge to bioanalytical chemists because of the following reasons: The products of oxidative protein damage are chemically diverse; protein oxidation generally occurs at low background levels; and the complexity of biological samples introduces high background noise when standard techniques such as immunolabeling are applied to "dirty" tissue extracts containing endogenous immunoglobulins or small molecular weight, chemically reactive compounds has been developed which circumvents these difficulties by incorporating a biotin label at sites of protein carbonylation. Biotin Hydrazide-labeled proteins are detectable using standard streptavidin-coupled detection techniques such as peroxidase-catalyzed chemiluminescence of immunoblots. Advantages of the Biotin Hydrazide-labeling technique are its sensitivity and its lack of reliance upon antibodies that inevitably suffer from nonspecific background noise and contaminating endogenous immunoglobulins.

Detection of protein carbonyls by means of biotin hydrazide-streptavidin affinity methods.[Pubmed:19378083]

Methods Mol Biol. 2009;536:457-62.

Oxidative posttranslational protein modifications occur as a normal process of cell biology and to a greater extent during pathogenic conditions. The detection and quantitation of protein oxidation has posed a continuing challenge to bioanalytical chemists because the products of oxidative protein damage are chemically diverse, protein oxidation generally occurs at low background levels, and the complexity of biological samples introduces high background noise when standard techniques such as immunolabeling are applied to "dirty" tissue extracts. A refinement of classic reductive amination methods has been developed, which circumvents these difficulties by incorporating a biotin label at sites of protein carbonylation. Biotin Hydrazide-labeled proteins are detectable using standard streptavidin-coupled detection techniques such as peroxidase-catalyzed chemiluminescence of immunoblots. Advantages of the Biotin Hydrazide-labeling technique are its sensitivity and its lack of reliance upon antibodies that inevitably suffer from nonspecific background noise and contaminating endogenous immunoglobulins.