Complex assembly of calgranulins A and B, two S100-like calcium-binding proteins from pig granulocytes

DellAngelica, EC; Schleicher, CH; Simpson, RJ; Santome, JA

Author(s): DellAngelica, EC; Schleicher, CH; Simpson, RJ; Santome, JA;
Details: Publication Year 1996-01,Volume 28,Issue #1,Page 53-62
Journal Title: INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY
Publication Type: Journal Article
Abstract: Calgranulin A (CAGA) and calgranulin B (CAGB) are two S100-like calcium-binding proteins that in human, bovine and mouse granulocytes are associated into a heterocomplex. We have previously identified in pig granulocytes the porcine homologue of CAGA and a novel S100-like protein which was named calgranulin C (CAGC). As pig CAGA is not associated with CAGC, we herein investigate its possible association with other proteins. CAGA was purified from pig granulocytes by gel filtration followed by Mono Q chromatography. The purified fractions were analysed by SDS-polyacrylamide gel electrophoresis, isoelectric focusing, mass spectrometry, chemical cross-linking and hydrophobic interaction chromatography. The CAGA-associated protein was further characterized by amino acid sequencing. Two CAGA-containing fractions were isolated. One of them was identified as a CAGA homodimer. The other fraction consists of a heterocomplex containing CAGA and a pI 7.0 calcium-binding protein; this protein has a molecular mass of 15,877.9 +/- 3.8 Da (mean +/- SD) whereas it migrates on 10 and 16% polyacrylamide gels as a 24- and 20-kDa protein, respectively. The pI 7.0 protein was identified by internal amino acid sequencing as the porcine homologue of CAGE. The stoichiometry of the heterocomplex was estimated to be 1:1. Both the CAGA homodimer and CAGA/CAGB were found to be non-covalently associated. Unlike the homodimer, CAGA/CAGB was bound to a Phenyl Superose column in a calcium-dependent manner. Our results suggest that pig granulocytes contain, in addition to CAGC, a CAGA homodimer and a CAGA/CAGB heterodimer. It is proposed that CAGA/CAGB and the CAGA homodimer may play different roles in vivo.
Publisher: PERGAMON-ELSEVIER SCIENCE LTD
Keywords: GEL-ELECTROPHORESIS; CONFORMATIONAL-CHANGES; S100 PROTEINS; KINASE-C; DIFFERENTIATION; SEQUENCE; MRP14; PURIFICATION; EXPRESSION; MEMBRANE
Publisher's Version: https://doi.org/10.1016/1357-2725(95)00115-8
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 1996-01-01 12:00:00