Production of human skeletal α-actin proteins by the baculovirus expression system

P.A. Akkari; K.J. Nowak; K. Beckman; K.R. Walker; F. Schachat; N.G Laing

doi:10.1016/S0006-291X(03)01133-1

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Production of human skeletal α-actin proteins by the baculovirus expression system

Journal article

Peer reviewed

Production of human skeletal α-actin proteins by the baculovirus expression system

P.A. Akkari, K.J. Nowak, K. Beckman, K.R. Walker, F. Schachat and N.G Laing

Biochemical and Biophysical Research Communications, Vol.307(1), pp.74-79

2003

DOI: https://doi.org/10.1016/S0006-291X(03)01133-1

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Abstract

Mutations within the human skeletal muscle α-actin gene cause three different skeletal muscle diseases. Functional studies of the mutant proteins are necessary to better understand the pathogenesis of these diseases, however, no satisfactory system for the expression of mutant muscle actin proteins has been available. We investigated the baculovirus expression vector system (BEVS) for the abundant production of both normal and mutant skeletal muscle α-actin. We show that non-mutated actin produced in the BEVS behaves similarly to native actin, as shown by DNase I affinity purification, Western blotting, and consecutive cycles of polymerisation and depolymerisation. Additionally, we demonstrate the production of mutant actin proteins in the BEVS, without detriment to the insect cells in which they are expressed. The BEVS therefore is the method of choice for studying mutant actin proteins causing human diseases.

Details

Title: Production of human skeletal α-actin proteins by the baculovirus expression system
Authors/Creators: P.A. Akkari (Author/Creator)
K.J. Nowak (Author/Creator)
K. Beckman (Author/Creator)
K.R. Walker (Author/Creator)
F. Schachat (Author/Creator)
N.G Laing (Author/Creator)
Publication Details: Biochemical and Biophysical Research Communications, Vol.307(1), pp.74-79
Publisher: Academic Press
Identifiers: 991005541138407891
Copyright: © 2003 Elsevier Science (USA).
Murdoch Affiliation: School of Veterinary and Biomedical Sciences
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.96 Cell Biology; 1.96.492 Myosin
Web Of Science research areas: Biochemistry & Molecular Biology; Biophysics
ESI research areas: Biology & Biochemistry