𝔖 Bobbio Scriptorium
✦   LIBER   ✦

Cysteine-Specific Radioiodination of Proteins with Fluorescein Maleimide

✍ Scribed by Michael Palmer; Martin Buchkremer; Angela Valeva; Sucharit Bhakdi

Publisher
Elsevier Science
Year
1997
Tongue
English
Weight
120 KB
Volume
253
Category
Article
ISSN
0003-2697

No coin nor oath required. For personal study only.

✦ Synopsis

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.

πŸ“œ SIMILAR VOLUMES

Incorporation of specifically labeled cy
Incorporation of specifically labeled cysteine into Escherichia coli protein
✍ Hal S. Beilan; George L. Kenyon; Douglas T. Browne πŸ“‚ Article πŸ“… 1983 πŸ› Elsevier Science 🌐 English βš– 445 KB

Protein obtained from several strains of Escherichia coli grown in the presence of [3,3'-14C]cystine contained the radiolabel in nearly all the other amino acids, suggesting catabolism of cysteine to pyruvic acid. Utilization in amino acid synthesis of the pyruvate thus generated can be blocked by g

Cysteine-Specific, Covalent Anchoring of
Cysteine-Specific, Covalent Anchoring of Transition Organometallic Complexes to the Protein Papain from Carica papaya
✍ Pierre Haquette; MichΓ¨le Salmain; Karolina Svedlung; Annie Martel; Bogna Rudolf; πŸ“‚ Article πŸ“… 2007 πŸ› John Wiley and Sons 🌐 English βš– 161 KB

## Abstract Site‐directed and covalent introduction of various transition metal–organic entities to the active site of the cysteine endoproteinase, papain, was achieved by treatment of this enzyme with a series of organometallic maleimide derivatives specially designed for the purpose. Kinetic stud

Identification of Cysteine Residues Alky
Identification of Cysteine Residues Alkylated with 3-Bromopropylamine by Protein Sequence Analysis
✍ R.A. Jue; J.E. Hale πŸ“‚ Article πŸ“… 1993 πŸ› Elsevier Science 🌐 English βš– 479 KB

A new reagent for the routine identification of cysteine residues during protein sequencing is described. This method employs 3-bromopropylamine to alkylate cysteines in proteins after reduction with dithiothreitol. Upon sequencing of the protein on an Applied Biosystems 477 A protein sequencer, the