RRC ID 6671
Author Ohkouchi S, Nishio K, Maeda M, Hitomi K, Adachi H, Maki M.
Title Identification and characterization of two penta-EF-hand Ca(2+)-binding proteins in Dictyostelium discoideum.
Journal J Biochem
Abstract Penta-EF-hand (PEF) proteins such as ALG-2 (apoptosis-linked gene 2 product) and the calpain small subunit are a newly classified family of Ca(2+)-binding proteins that possess five EF-hand-like motifs. We identified two mutually homologous PEF proteins, designated DdPEF-1 and DdPEF-2 (64% amino acid residue identities), in the cellular slime mold Dictyostelium discoideum. Both PEF proteins showed a higher similarity to mammalian ALG-2 and peflin (Group I PEF proteins) than to calpain and sorcin subfamily (Group II PEF proteins) in the first EF-hand (EF-1) regions. Northern blot analyses revealed that DdPEF-1 and DdPEF-2 were constitutively expressed throughout development of Dictyostelium, but their levels of expression were developmentally regulated. In situ hybridization analyses demonstrated that DdPEF-1 was expressed in both the anterior prestalk and the posterior prespore regions of the tipped aggregate, slugs and early culminants. On the other hand, DdPEF-2 was dominantly expressed in the anterior tip region of these multicellular structures. Both PEF proteins were detected as 22-23-kDa proteins in soluble fractions in the presence of EGTA but in particulate fractions in the presence of Ca(2+) by Western blotting using specific monoclonal antibodies. Together with the finding of PEF-like sequences in DNA databases of plants, fungi and protists, our results strongly suggest that Group I PEF proteins are ubiquitously present in all eukaryotes and play important roles in basic cellular functions.
Volume 130(2)
Pages 207-15
Published 2001-8-1
DOI 10.1093/oxfordjournals.jbchem.a002974
PMID 11481037
MeSH Amino Acid Sequence Animals Calcium / metabolism Calcium-Binding Proteins / chemistry* Calcium-Binding Proteins / classification Calcium-Binding Proteins / genetics Calcium-Binding Proteins / metabolism* Cell Fractionation Dictyostelium / chemistry* Dictyostelium / genetics EF Hand Motifs* Gene Expression Regulation, Developmental In Situ Hybridization Molecular Sequence Data Phylogeny Protozoan Proteins / chemistry Protozoan Proteins / classification Protozoan Proteins / genetics Protozoan Proteins / metabolism Sequence Alignment
IF 2.476
Times Cited 21
Cellular slime molds G00626 G01245