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Updated: Jul 20, 2026

Quantifying the Binding Interactions Between Cu(II) and Peptide Residues in the Presence and Absence of Chromophores
Published on: April 5, 2022
Riboflavin binding protein contains a type II copper binding site
Sheila R Smith1, Irina Pala, Marilee Benore-Parsons
1Department of Natural Sciences, University of Michigan - Dearborn, 4901 Evergreen Road, Dearborn, MI 48128, United States. sheilars@umd.umich.edu <sheilars@umd.umich.edu>
Egg white riboflavin binding protein binds copper(II) in a 1:1 ratio, suggesting a potential role in copper storage or transport. This protein may have a previously unrecognized function in handling copper ions.
Area of Science:
- Biochemistry
- Biometals
- Protein Chemistry
Background:
- Riboflavin binding protein (RBP) is primarily known for its role in vitamin B2 transport.
- The presence of metal ions in egg white proteins is not extensively characterized.
- Understanding RBP's interactions with other molecules can reveal novel biological functions.
Purpose of the Study:
- To investigate the potential interaction between purified egg white riboflavin binding protein and copper(II) ions.
- To characterize the binding stoichiometry and site characteristics of copper bound to RBP.
- To explore a potential novel function of RBP in copper homeostasis.
Main Methods:
- Purification of riboflavin binding protein from egg white.
- Dialysis experiments to assess copper(II) binding.
- Molar ratio determination of protein to copper.
- Electron paramagnetic resonance (EPR) spectroscopy to identify copper binding sites.
Main Results:
- Riboflavin binding protein binds copper(II) in an approximate 1:1 molar ratio under dialysis conditions.
- A small but significant amount of copper is inherently present in the purified RBP.
- Electron paramagnetic resonance (EPR) spectroscopy identified a single type II copper binding site.
Conclusions:
- Riboflavin binding protein possesses the ability to bind copper(II) ions.
- The presence of an intrinsic copper site suggests a possible role in copper metabolism.
- These findings propose a previously unrecognized function for RBP in copper storage or transport.
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