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Sequestered actin in chick embryo fibroblasts

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Abstract

Chick embryo fibroblasts contain about 75-100 μM unpolymerized actin and at least four proteins which can bind actin monomers, actin depolymerizing factor (ADF), gelsolin, profilin, and thymosin β4 (Tβ4). Fibroblast extracts are analyzed by non-denaturing polyacrylamide gel electrophoresis and immunoblotting where most of the G-actin is detected as a complex with Tβ4. When fibroblast extracts are fractionated by gel filtration and the fractions are analyzed by PAGE and HPLC, most of the G-actin elutes in a peak that also contains Tβ4 at an overall molar ratio of 1.9:1 relative to actin. Gelsolin, profilin, and ADF are also detectable in the gel filtration eluate and at least partly coelute with actin, and account for only a minor fraction of the soluble actin pool. These observations indicate that under the growth conditions studied, Tβ4 is the major actin-sequestering protein in fibroblasts.

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, University of Pennsylvania, Philadelphia, PA, 19104-6058, USA

    K. Nagamalleswari & Daniel Safer

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  1. K. Nagamalleswari
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  2. Daniel Safer
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Nagamalleswari, K., Safer, D. Sequestered actin in chick embryo fibroblasts. Mol Cell Biochem 209, 63–67 (2000). https://doi.org/10.1023/A:1007025602110

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