K. Duus, N. Sandhu, C. S. Jorgensen, P. R. Hansen, A. Steino, M. Thaysen-Andersen, P. Hojrup and G. Houen Pages 1414 - 1423 ( 10 )
The interaction of calreticulin with native and denatured forms and polypeptides in proteolytic digests of proteins representing structural classes of all-α-helix (hemoglobin, serum albumin), all-β-sheet (IgG) and α-helix + β-sheets (lysozyme, ovalbumin) was investigated. The binding of calreticulin to denatured proteins was found to depend on conformation and structural class of the protein. No interaction was observed with the native proteins, whereas binding was seen for the denatured proteins, the order of interaction being lysozyme = IgG > ovalbumin hemoglobin = serum albumin. Moreover, the interaction between calreticulin and the heat-denatured proteins depended on the temperature and time used for denaturation and the degree of proteolytic fragmentation. Calreticulin bound well to peptides in proteolytic digests from protease K or chymotrypsin treatment of lysozyme, IgG and ovalbumin but weakly or not at all to peptides in proteolytic digests of hemoglobin and serum albumin. Synthetic peptides from lysozyme and ovalbumin confirmed binding to hydrophobic peptides from these proteins. These results show that calreticulin has the ability to interact with denatured and fragmented forms of proteins with a preference for β-strand structure and hydrophobicity.
Calreticulin, peptide specificity, chaperone, structural class, α-helix, β-sheet
Department of Clinical Biochemistry and Immunology, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen, Denmark.