Some properties of purified skeletal muscle α actinin

A. Suzuki, D. E. Goll, I. Singh, R. E. Allen, R. M. Robson, M. H. Stromer

Research output: Contribution to journalArticle

137 Scopus citations

Abstract

Highly purified α actinin can be made by using the low ionic strength extraction procedure and then subjecting the crude α actinin fraction obtained with this extraction procedure to successive chromatography on DEAE cellulose and hydroxyapatite. Hydroxyapatite chromatography specifically removes a protein having a subunit molecular weight of 42,000 on sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Hydroxyapatite purified α actinin sediments entirely as a 6.21 S boundary in the analytical ultracentrifuge with no trace of the small 9 to 10 S boundary seen in earlier α actinin preparations purified by DEAE cellulose chromatography. In 100 mM KCl, 20 mM Tris.acetate, pH 7.5, hydroxyapatite purified α actinin has a diffusion coefficient (Do 20,w) of 2.71 x 10 -7 cm 2.s -1, an intrinsic viscosity of 20.6 ml.g -1, a molecular weight of 201,000 ± 4,300 (plus or minus least squares standard error) as determined by sedimentation equilibrium, and a molecular weight of 210,000 as determined by sedimentation diffusion. In 6 M guanidine HCl, hydroxyapatite purified α actinin has a molecular weight of 106,000 ± 6,300 as determined by sedimentation equilibrium and a molecular weight of 100,000 as determined by a calibrated 4% agarose gel permeation column. SDS polyacrylamide gel electrophoresis gives a molecular weight of 96,000 to 100,000 for hydroxyapatite purified α actinin. Rod shaped particles 44 x 390 to 400 Å are seen in electron micrographs of negatively stained α actinin. By assuming 45% hydration and a molecular weight of 206,000, dimensions of approximately 40 x 500Å can be calculated for the α actinin molecule by using either So 20,w, Do 20,w, intrinsic viscosity, or a calibrated 6% agarose gel permeation column. Hydroxyapatite purified α actinin has an α helical content of 74% as measured by circular dichroism at 208 nm.

Original languageEnglish (US)
Pages (from-to)6860-6870
Number of pages11
JournalJournal of Biological Chemistry
Volume251
Issue number21
StatePublished - Dec 1 1976
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Suzuki, A., Goll, D. E., Singh, I., Allen, R. E., Robson, R. M., & Stromer, M. H. (1976). Some properties of purified skeletal muscle α actinin. Journal of Biological Chemistry, 251(21), 6860-6870.