Cysteine-Specific Radioiodination of Proteins with Fluorescein Maleimide

6533b839fe1ef96bd12a5b9a

RESEARCH PRODUCT

Cysteine-Specific Radioiodination of Proteins with Fluorescein Maleimide

Martin Buchkremer Sucharit Bhakdi Angela Valeva Michael Palmer

subject

Erythrocytes Polymers Thiosulfates Biophysics Plasma protein binding Sodium thiosulfate Complement Hemolytic Activity Assay Sensitivity and Specificity Biochemistry Iodine Radioisotopes Tosyl Compounds chemistry.chemical_compound Bacterial Proteins Cysteine Fluorescein Molecular Biology Chloramine Chromatography Chloramines Proteins Halogenation Cell Biology Fluoresceins Biochemistry chemistry Spectrophotometry Reagent Streptolysins Chromatography Gel Streptolysin Protein Binding Cysteine

description

A protocol is described for coupling of carrier-free iodine to protein sulfhydryl groups via fluorescein maleimide. 125I is first coupled to fluorescein maleimide in the presence of chloramine T. Iodination is stopped with sodium thiosulfate, and the iodine-substituted fluorescein maleimide is reacted with free cysteines of the protein. Excess label is then removed by gel-permeation chromatography. The procedure avoids exposition of the protein to oxidative conditions and does not require purification of the labeled carrier reagent. Suitability of the method for a given protein can be evaluated spectrophotometrically without employing radioactivity. It can be applied under denaturing conditions and may be particularly useful with mutant proteins carrying engineered single cysteine residues at sites that are not functionally critical.

year	journal	country	edition	language
1997-12-31	Analytical Biochemistry

https://doi.org/10.1006/abio.1997.2364

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