Calcium ion binding to delta- and to beta-crystallins. The presence of the "EF-hand" motif in delta-crystallin that aids in calcium ion binding.
Abnormal levels of endogenous calcium ions are known to induce eye lens opacity, and a variety of causative factors has been proposed, including calcium-mediated aggregation and precipitation of the lens proteins crystallins. We have specifically looked in some detail at the interaction of Ca2+ with various crystallins and its consequences. Lenses incubated in solutions containing 10 mM Ca2+ or 5 mM Tb3+ opacified. Fluorescence titration of crystallins with TbCl3 revealed that this ion binds to delta- and beta-crystallins in solution. Equilibrium dialysis showed that four Ca2+ ions bind to one delta-crystallin tetramer with an affinity of 4.3 x 10(3) M-1. Analysis of the amino acid sequence of delta-crystallin reveals the presence of a calmodulin-type "helix-loop-helix" or "EF-hand" calcium ion binding conformational motif in the region comprising residues 300-350. This is a novel feature of the molecule not reported so far. No other crystallins appear to have this motif. beta-Crystallin also binds four Ca2+ ions/aggregate unit of mass 160 kDa, with an affinity of 2.6 x 10(3) M-1, presumably in the midregion of the molecule that is rich in anionic and polar residues. Circular dichroism spectroscopy shows that the binding of calcium ion leads to subtle conformational changes in the molecules, notably in the tertiary structure.
Sharma
Y
Y
Centre for Cellular and Molecular Biology, Hyderabad, India.
Rao
C M
CM
Narasu
M L
ML
Rao
S C
SC
Somasundaram
T
T
Gopalakrishna
A
A
Balasubramanian
D
D
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002122:Calcium Chloride; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D002645:Chickens; D002942:Circular Dichroism; D003459:Crystallins / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology; D007811:Lanthanum; D008969:Molecular Sequence Data; D010445:Peptide Elongation Factors / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D011487:Protein Conformation; D051381:Rats; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D002135:Calcium-Binding Proteins; D003459:Crystallins; D010445:Peptide Elongation Factors; C028521:lanthanum chloride; D007811:Lanthanum; D002122:Calcium Chloride; D002118:Calcium