Sequential Calcium Binding to the Regulatory Domain of Calcium Vector Protein Reveals Functional Asymmetry and a Novel Mode of Structural Rearrangement
Autor: | Sibyl Baladi, Isabelle Théret, Hiroshi Sakamoto, Jos A. Cox, Constantin T. Craescu |
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Rok vydání: | 2000 |
Předmět: |
Magnetic Resonance Spectroscopy
Sequence Homology Amino Acid Calmodulin biology Protein Conformation Molecular Sequence Data Muscle Proteins chemistry.chemical_element Nuclear magnetic resonance spectroscopy Calcium Biochemistry Recombinant Proteins Molten globule Protein tertiary structure Crystallography Ion binding chemistry biology.protein Amino Acid Sequence Protein secondary structure Binding domain |
Zdroj: | Biochemistry. 39:7920-7926 |
ISSN: | 1520-4995 0006-2960 |
DOI: | 10.1021/bi000360z |
Popis: | Calcium vector protein (CaVP) from amphioxus is a two-domain, calcium-binding protein (18.3 kDa) of the calmodulin superfamily. Only two of the four EF-hand motifs (sites III and IV) have a significant binding affinity for calcium ions. We determined the solution structure of the domain containing these active sites (C-CaVP: W81-S161), in the Ca(2+)-saturated state, using NMR spectroscopy and restrained molecular dynamics. The tertiary structure is similar to other Ca(2+)-binding domains containing a pair of EF-hand motifs. The apo state has spectroscopic and thermodynamic characteristics of a molten globule, with conserved secondary structure but highly fluctuating tertiary organization. Titration of C-CaVP with Ca(2+) revealed a stepwise ion binding, with a stable equilibrium intermediate in which only site III binds a calcium ion. Despite a highly fluctuating structure of the free site IV, the calcium-bound site III has a persistent structure, with similar secondary elements but different interhelix angle and hydrophobic packing relative to the fully calcium-saturated state. |
Databáze: | OpenAIRE |
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