Small espin: a third actin-bundling protein and potential forked protein ortholog in brush border microvilli.
An approximately 30-kD isoform of the actin-binding/ bundling protein espin has been discovered in the brush borders of absorptive epithelial cells in rat intestine and kidney. Small espin is identical in sequence to the COOH terminus of the larger ( approximately 110-kD) espin isoform identified in the actin bundles of Sertoli cell-spermatid junctional plaques (Bartles, J.R., A. Wierda, and L. Zheng. 1996. J. Cell Sci. 109:1229-1239), but it contains two unique peptides at its NH2 terminus. Small espin was localized to the parallel actin bundles of brush border microvilli, resisted extraction with Triton X-100, and accumulated in the brush border during enterocyte differentiation/migration along the crypt-villus axis in adults. In transfected BHK fibroblasts, green fluorescent protein-small espin decorated F-actin-containing fibers and appeared to elicit their accumulation and/or bundling. Recombinant small espin bound to skeletal muscle and nonmuscle F-actin with high affinity (Kd = 150 and 50 nM) and cross-linked the filaments into bundles. Sedimentation, gel filtration, and circular dichroism analyses suggested that recombinant small espin was a monomer with an asymmetrical shape and a high percentage of alpha-helix. Deletion mutagenesis suggested that small espin contained two actin-binding sites in its COOH-terminal 116-amino acid peptide and that the NH2-terminal half of its forked homology peptide was necessary for bundling activity.
Bartles
J R
JR
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA. j-bartles@nwu.edu
Zheng
L
L
Li
A
A
Wierda
A
A
Chen
B
B
D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D006224:Cricetinae; D015723:Gene Library; D049452:Green Fluorescent Proteins; D007413:Intestinal Mucosa / Q000648:ultrastructure; D007668:Kidney / Q000378:metabolism / Q000648:ultrastructure; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D008871:Microvilli / Q000378:metabolism* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017433:Protein Structure, Secondary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014162:Transfection
D000199:Actins; D008164:Luminescent Proteins; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; C104108:espin protein, mouse; D049452:Green Fluorescent Proteins