title: Characterization of a calsequestrin-like protein from sea-urchin eggs.
abstract: Following our studies on the identification of a calsequestrin-like protein (CSLP) from sea-urchin eggs [Oberdorf, Lebeche, Head & Kaminer (1988) J. Biol Chem. 263, 6806-6809], we have characterized its Ca(2+)-binding properties and identified it as a glycoprotein. The molecule binds 23 mol of Ca2+/mol of protein, as determined by equilibrium dialysis. This is in the range reported for cardiac calsequestrin but is about half the binding capacity of striated muscle calsequestrin. The affinities of the CSLP for Ca2+ are decreased by increasing KCl concentrations (20-250 mM) and the presence of Mg2+ (3 mM) in the medium: the half-maximal binding values varied from 1.62 to 5.77 mM. Hill coefficients indicated mild co-operativity in the Ca2+ binding. Ca2+ (1-8 mM)-induced u.v. difference spectra and intrinsic fluorescence changes suggest a net exposure of aromatic residues to an aqueous environment. C.d. measurements showed minor Ca(2+)-induced changes in alpha-helical and beta-sheet content of less than 10%. These spectral changes are distinctly different from those found in muscle calsequestrin. Immunoblotting studies showed that the CSLP is distinct from calreticulin, a low-affinity Ca(2+)-binding protein.
authors.affiliation: Department of Physiology, Boston University School of Medicine, MA 02118.
authors.forename: D
authors.initials: D
authors.lastname: Lebeche
authors.forename: B
authors.initials: B
authors.lastname: Kaminer
mesh_terms: D000595:Amino Acid Sequence; D000606:Amino Sugars / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002155:Calsequestrin / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D011487:Protein Conformation; D012617:Sea Urchins; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000606:Amino Sugars; D002155:Calsequestrin; D006023:Glycoproteins; D002118:Calcium