Journal of Muscle Research & Cell Motility

, Volume 19, Issue 6, pp 639–646 | Cite as

Generation of functional β-actinin (CapZ) in an E. coli expression system

  • Yoshinori Soeno
  • Hiroshi Abe
  • Sumiko Kimura
  • Koscak Maruyama
  • Takashi Obinata
Article

Abstract

β-actinin (CapZ) is a heterodimeric actin-binding protein which caps the barbed end of actin filaments and nucleates actin-polymerization in a Ca2+-independent manner. In myofibrils it is localized in the Z-lines. As judged by these properties of β-actinin, it is conceivable that β-actinin is involved in the regulation of actin assembly, especially in the formation of I-Z-I complex during myofibrillogenesis. In this study, we devised a system to produce functional β-actinin in E. Coli.The cDNAs of βI′ and βII subunits of β-actinin were obtained by RT-PCR methods using the published sequence as references, and subcloned in a pET vector. When the proteins were produced with the cDNA of either βI′ or βII in E. coli, the proteins were insoluble and non-functional. However, when the cDNAs encoding the two subunits were cloned into a single vector and␣both proteins were expressed simultaneously, the proteins became soluble and purified as a functional heterodimer. The␣activity of the purified proteins was not distinguishable from that of β-actinin purified from skeletal muscle.

Keywords

Skeletal Muscle Expression System Actin Filament Barb Independent Manner 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Yoshinori Soeno
  • Hiroshi Abe
  • Sumiko Kimura
  • Koscak Maruyama
  • Takashi Obinata

There are no affiliations available

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