Calcium-induced human centrin 1 self-assembly and double-regulating the binding with peptide R18-Sfi1p.

Centrin is a member of the EF-hand super-family that plays pivotal role in the centrosome duplication and separation. In the present paper, we characterized the properties of metal ions as well as peptide R18-Sfi1p binding to human centrin 1 (HsCen1) by fluorescence spectra and isothermal titration calorimetry (ITC). Four metal ions binding sites on HsCen1 were identified through ITC experiments. The conditional binding constants of the EF-hand domain on HsCen1 with Ca2+ were quantitatively calculated. In reversible manner, Ca2+ can induce HsCen1 self-assembly. In addition, HsCen1 bound with peptide R18-Sfi1p in calcium-dependent with middle-affinity. Phosphorylation at Ser170 weakened interaction HsCen1 with the substrate and removal calcium ions further weakened interactions of the two molecules. Hence, we inferred that centrin initiating downstream peptides may be a double-regulated process by calcium and phosphorylation. These results are of significance for understanding the relationship between PTM and metal regulation.