A novel isoform of cytoplasmic actin that binds poly-L-proline.

Abstract

An actin-like protein was purified to apparent homogeneity from chick-embryo homogenates and chick-embryo fibroblasts by the use of poly-L-proline-agarose affinity chromatography; we therefore refer to this protein as PBP (poly-L-proline-binding protein). PBP binds to deoxyribonuclease-agarose, co-migrates with known actin standards on SDS/polyacrylamide-gel electrophoresis, and has an amino acid composition similar to that of actin. Linear peptide maps after digestion with Staphylococcus aureus proteinase reveal its apparent homology with gamma-actin; however, isoelectric-focusing experiments show that PBP is clearly more acidic than any of the three major isoforms of actin. PBP polymerizes in the presence of ATP to form fibrillar structures resembling actin paracrystalline aggregates. In chick-embryo fibroblasts, immunofluorescence with antibodies to PBP shows that its distribution is cytoplasmic: perinuclear staining of the cytoplasm, generalized cytoplasmic staining and peripheral fibrillar structures are evident. In contrast, antibodies specific for the (alpha, gamma)-actins reveal the typical stress fibre structures characteristic of fibroblastic cells. PBP appears to constitute a novel isoform of cellular actin, distinct from the known actin isoforms in terms of its lower isoelectric point, its ability to bind poly-L-proline and its distinct subcellular localization.