The 23-kilodalton protein, a substrate of protein kinase C, in bovine neutrophil cytosol is a member of the S100 family.
A bovine neutrophil protein termed p23 because of an apparent molecular mass of 23 kDa in SDS-PAGE is present in large amounts both in a soluble form in the cytosolic fraction of bovine neutrophil homogenates and associated to the cytoskeleton. P23 is accompanied during the first steps of the purification procedure by a smaller size protein termed p7 on the basis of a rate of migration in SDS-PAGE corresponding to a 7-kDa protein [Stasia, M. J., Dianoux, A. C., & Vignais, P. V. (1989) Biochemistry 28, 9659-9667]. The two proteins, p23 and p7, have been purified to homogeneity by an improved procedure consisting of two chromatographic steps. The electrospray mass spectrometry technique applied to p23 and p7 indicated molecular masses close to 17 and 10 kDa, respectively, significantly different from the masses derived by SDS-PAGE. Bovine neutrophil p23 and p7 presented large primary structure homologies with two human proteins, MRP14 and MRP8, which are expressed in large amounts in macrophages under conditions of chronic inflammation. In addition, p23 and p7 cross-reacted with monoclonal antibodies specific of MRP14 and MRP8. Bovine p23 and p7 bound Ca2+, and their amino acid sequences contained two Ca(2+)-binding domains per protein, largely identical to those of human MRP14 and MRP8. Bovine p23 and p7 associated together to form a heterodimeric complex, which largely escaped attack by trypsin, whereas the isolated p23 and p7 components were readily digested. These features are typical of Ca(2+)-binding proteins belonging to the S100 family.(ABSTRACT TRUNCATED AT 250 WORDS)
Dianoux
A C
AC
Département de Biologie Moléculaire et Structurale, Centre d'Etudes Nucléaires, Grenoble, France.
Stasia
M J
MJ
Garin
J
J
Gagnon
J
J
Vignais
P V
PV
D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000737:chemistry / Q000378:metabolism; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D040501:Calgranulin A; D040502:Calgranulin B; D002417:Cattle; D003488:Cyanogen Bromide; D003594:Cytoplasmic Granules / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry; D010587:Phagocytosis; D011487:Protein Conformation; D011493:Protein Kinase C / Q000378:metabolism*; D009418:S100 Proteins / Q000737:chemistry*; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D000943:Antigens, Differentiation; D002135:Calcium-Binding Proteins; D040501:Calgranulin A; D040502:Calgranulin B; D046911:Macromolecular Substances; D010446:Peptide Fragments; D009418:S100 Proteins; C075304:calgranulin B, bovine; D011493:Protein Kinase C; D003488:Cyanogen Bromide; D002118:Calcium