4YI9

Crystal structure of non-myristoylated E153A recoverin at 1.35 A resolution with a sodium ion bound to EF-hand 2 and calcium ion bound to EF-hand 3

Summary for 4YI9

• Entry DOI: 10.2210/pdb4yi9/pdb
• Related: 4MLW 4YI8
• Descriptor: Recoverin, SODIUM ION, CALCIUM ION, ... (4 entities in total)
• Functional Keywords: calcium binding protein, ef hand, ncs family
• Biological source: Bos taurus (Bovine)
• Total number of polymer chains: 1
• Total formula weight: 23240.24

Authors

Prem Kumar, R.,Ranaghan, M.J.,Oprian, D.D. (deposition date: 2015-02-28, release date: 2015-12-02, Last modification date: 2023-09-27)

Primary citation

Kumar, R.P.,Ranaghan, M.J.,Ganjei, A.Y.,Oprian, D.D.
Crystal Structure of Recoverin with Calcium Ions Bound to Both Functional EF Hands.
Biochemistry, 54:7222-7228, 2015

PubMed Abstract: Recoverin (Rv), a small Ca(2+)-binding protein that inhibits rhodopsin kinase (RK), has four EF hands, two of which are functional (EF2 and EF3). Activation requires Ca(2+) in both EF hands, but crystal structures have never been observed with Ca(2+) ions in both sites; all previous structures have Ca(2+) bound to only EF3. We suspected that this was due to an intermolecular crystal contact between T80 and a surface glutamate (E153) that precluded coordination of a Ca(2+) ion in EF2. We constructed the E153A mutant, determined its X-ray crystal structure to 1.2 Å resolution, and showed that two Ca(2+) ions are bound, one in EF3 and one in EF2. Additionally, several other residues are shown to adopt conformations in the 2Ca(2+) structure not seen previously and not seen in a second structure of the E153A mutant containing Na(+) instead of Ca(2+) in the EF2 site. The side-chain rearrangements in these residues form a 28 Å allosteric cascade along the surface of the protein connecting the Ca(2+)-binding site of EF2 with the active-site pocket responsible for binding RK.

PubMed: 26584024
DOI: 10.1021/acs.biochem.5b01160

Experimental method

X-RAY DIFFRACTION (1.35 Å)