New fluorescent reagents specific for Ca{sup 2+}-binding proteins
- Department of Life Sciences and the NIBN, Ben-Gurion University, Beer-Sheva 84105 (Israel)
- Department of Physiology, Ben-Gurion University, Beer-Sheva 84105 (Israel)
Highlights: Black-Right-Pointing-Pointer New reagents specifically inhibit the activity of Ca{sup 2+}-dependent proteins. Black-Right-Pointing-Pointer FITC-Ru and EITC-Ru allow for mechanism-independent probing of Ca{sup 2+}-binding proteins. Black-Right-Pointing-Pointer Changes in reagents fluorescence allow characterization of protein Ca{sup 2+}-binding properties. -- Abstract: Ca{sup 2+} carries information pivotal to cell life and death via its interactions with specific binding sites in a protein. We previously developed a novel photoreactive reagent, azido ruthenium (AzRu), which strongly inhibits Ca{sup 2+}-dependent activities. Here, we synthesized new fluorescent ruthenium-based reagents containing FITC or EITC, FITC-Ru and EITC-Ru. These reagents were purified, characterized and found to specifically interact with and markedly inhibit Ca{sup 2+}-dependent activities but not the activity of Ca{sup 2+}-independent reactions. In contrast to many reagents that serve as probes for Ca{sup 2+}, FITC-Ru and EITC-Ru are the first fluorescent divalent cation analogs to be synthesized and characterized that specifically bind to Ca{sup 2+}-binding proteins and inhibit their activity. Such reagents will assist in characterizing Ca{sup 2+}-binding proteins, thereby facilitating better understanding of the function of Ca{sup 2+} as a key bio-regulator.
- OSTI ID:
- 22210251
- Journal Information:
- Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 1 Vol. 426; ISSN 0006-291X; ISSN BBRCA9
- Country of Publication:
- United States
- Language:
- English
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