Journal article
Milligram quantities of homogeneous recombinant full-length mouse Munc18c from escherichia coli cultures
A Rehman, RJ Jarrott, AE Whitten, GJ King, SH Hu, MP Christie, BM Collins, JL Martin
Plos One | PUBLIC LIBRARY SCIENCE | Published : 2013
Abstract
Vesicle fusion is an indispensable cellular process required for eukaryotic cargo delivery. The Sec/Munc18 protein Munc18c is essential for insulin-regulated trafficking of glucose transporter4 (GLUT4) vesicles to the cell surface in muscle and adipose tissue. Previously, our biophysical and structural studies have used Munc18c expressed in SF9 insect cells. However to maximize efficiency, minimize cost and negate any possible effects of post-translational modifications of Munc18c, we investigated the use of Escherichia coli as an expression host for Munc18c. We were encouraged by previous reports describing Munc18c production in E. coli cultures for use in in vitro fusion assay, pulldown as..
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Awarded by National Health and Medical Research Council
Funding Acknowledgements
This research was supported by the National Health and Medical Research Council of Australia (NHMRC) program grant 535921 to JLM. AEW is an NHMRC Peter Doherty Fellow (569864), BMC is an Australian Research Council (ARC) Future Fellow (FT100100027) and JLM is an ARC Australian Laureate Fellow (FL0992138) and Honorary NHMRC Fellow. The IMB Mass Spectrometry Facility used in this research was funded in part through an ARC LIEF grant (LE110100186). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.