Nexilin: a novel actin filament-binding protein localized at cell–matrix adherens junction

T Ohtsuka, H Nakanishi, W Ikeda, A Satoh… - The Journal of cell …, 1998 - rupress.org
T Ohtsuka, H Nakanishi, W Ikeda, A Satoh, Y Momose, H Nishioka, Y Takai
The Journal of cell biology, 1998rupress.org
We isolated two novel actin filament (F-actin)–binding proteins from rat brain and rat 3Y1
fibroblast. They were splicing variants, and we named brain big one b-nexilin and fibroblast
small one s-nexilin. b-Nexilin purified from rat brain was a protein of 656 amino acids (aa)
with a calculated molecular weight of 78,392, whereas s-nexilin, encoded by the cDNA
isolated from rat 3Y1 cells by the reverse transcriptase-PCR method, was a protein of 606 aa
with a calculated molecular weight of 71,942. b-Nexilin had two F-actin–binding domains …
We isolated two novel actin filament (F-actin)–binding proteins from rat brain and rat 3Y1 fibroblast. They were splicing variants, and we named brain big one b-nexilin and fibroblast small one s-nexilin. b-Nexilin purified from rat brain was a protein of 656 amino acids (aa) with a calculated molecular weight of 78,392, whereas s-nexilin, encoded by the cDNA isolated from rat 3Y1 cells by the reverse transcriptase-PCR method, was a protein of 606 aa with a calculated molecular weight of 71,942. b-Nexilin had two F-actin– binding domains (ABDs) at the NH2-terminal and middle regions, whereas s-nexilin had one ABD at the middle region because 64 aa residues were deleted and 14 aa residues were inserted in the first NH2-terminal ABD of b-nexilin, and thereby the first ABD lost its activity. b- and s-nexilins bound along the sides of F-actin, but only b-nexilin showed F-actin cross-linking activity. b-Nexilin was mainly expressed in brain and testis, whereas s-nexilin was mainly expressed in testis, spleen, and fibroblasts, such as rat 3Y1 and mouse Swiss 3T3 cells, but neither b- nor s-nexilin was detected in liver, kidney, or cultured epithelial cells. An immunofluorescence microscopic study revealed that s-nexilin was colocalized with vinculin, talin, and paxillin at cell– matrix adherens junction (AJ) and focal contacts, but not at cell–cell AJ, in 3Y1 cells. Overexpressed b- and s-nexilins were localized at focal contacts but not at cell–cell AJ. These results indicate that nexilin is a novel F-actin–binding protein localized at cell–matrix AJ.
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