Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Marino, J; Bordag, N; Keller, S; Zerbe, O.
Mistic's membrane association and its assistance in overexpression of a human GPCR are independent processes.
Protein Sci. 2015; 24(1): 38-48.
Doi: 10.1002/pro.2582
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- Co-Autor*innen der Med Uni Graz
- Bordag Natalie
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- Abstract:
- The interaction of the Bacillus subtilis protein Mistic with the bacterial membrane and its role in promoting the overexpression of other membrane proteins are still matters of debate. In this study, we aimed to determine whether individual helical fragments of Mistic are sufficient for its interaction with membranes in vivo and in vitro. To this end, fragments encompassing each of Mistic's helical segments and combinations of them were produced as GFP-fusions, and their cellular localization was studied in Escherichia coli. Furthermore, peptides corresponding to the four helical fragments were synthesized by solid-phase peptide synthesis, and their ability to acquire secondary structure in a variety of lipids and detergents was studied by circular dichroism spectroscopy. Both types of experiments demonstrate that the third helical fragment of Mistic interacts only with LDAO micelles but does not partition into lipid bilayers. Interestingly, the other three helices interact with membranes in vivo and in vitro. Nevertheless, all of these short sequences can replace full-length Mistic as N-terminal fusions to achieve overexpression of a human G-protein-coupled receptor in E. coli, although with different effects on quantity and quality of the protein produced. A bioinformatic analysis of the Mistic family expanded the number of homologs from 4 to 20, including proteins outside the genus Bacillus. This information allowed us to discover a highly conserved Shine-Dalgarno sequence in the operon mstX-yugO that is important for downstream translation of the potassium ion channel yugO. © 2014 The Protein Society.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Bacillus subtilis - chemistry
- Bacillus subtilis - genetics
- Bacillus subtilis - metabolism
- Bacterial Proteins - chemistry
- Bacterial Proteins - genetics
- Bacterial Proteins - metabolism
- Cloning, Molecular - methods
- Escherichia coli - cytology
- Escherichia coli - genetics
- Escherichia coli - metabolism
- Green Fluorescent Proteins - chemistry
- Green Fluorescent Proteins - genetics
- Green Fluorescent Proteins - metabolism
- Humans -
- Lipid Bilayers - metabolism
- Micelles -
- Molecular Sequence Data -
- Peptides - chemistry
- Peptides - genetics
- Peptides - metabolism
- Protein Folding -
- Protein Structure, Secondary -
- Receptors, G-Protein-Coupled - chemistry
- Receptors, G-Protein-Coupled - genetics
- Receptors, G-Protein-Coupled - metabolism
- Recombinant Fusion Proteins - chemistry
- Recombinant Fusion Proteins - genetics
- Recombinant Fusion Proteins - metabolism
- Up-Regulation -
- Find related publications in this database (Keywords)
- Mistic
- GFP-fusion
- lipids
- peptide-membrane interaction
- yugO
- GPCR expression