Literature DB >> 33522420

Efficient expression of a cnidarian peptide-gated ion channel in mammalian cells.

Michèle Bachmann1, Audrey Ortega-Ramírez1, Lilia Leisle1, Stefan Gründer1.   

Abstract

Hydra Na+ channels (HyNaCs) are peptide-gated ion channels of the DEG/ENaC gene family that are directly activated by neuropeptides of the Hydra nervous system. They have previously been successfully characterized in Xenopus oocytes. To establish their expression in mammalian cells, we transiently expressed heteromeric HyNaC2/3/5 in human HEK 293 and monkey COS-7 cells. We found that the expression of HyNaC2/3/5 using native cDNAs was inefficient and that codon optimization strongly increased protein expression and current amplitude in patch-clamp experiments. We used the improved expression of codon-optimized channel subunits to perform Ca2+ imaging and to demonstrate their glycosylation pattern. In summary, we established efficient expression of a cnidarian ion channel in mammalian cell lines.

Entities:  

Keywords:  Codon optimization; Hydra; ligand-gated ion channel; neuropeptide

Mesh:

Substances:

Year:  2021        PMID: 33522420      PMCID: PMC7889164          DOI: 10.1080/19336950.2021.1882762

Source DB:  PubMed          Journal:  Channels (Austin)        ISSN: 1933-6950            Impact factor:   2.581


  38 in total

Review 1.  Back to the Basics: Cnidarians Start to Fire.

Authors:  Thomas C G Bosch; Alexander Klimovich; Tomislav Domazet-Lošo; Stefan Gründer; Thomas W Holstein; Gáspár Jékely; David J Miller; Andrea P Murillo-Rincon; Fabian Rentzsch; Gemma S Richards; Katja Schröder; Ulrich Technau; Rafael Yuste
Journal:  Trends Neurosci       Date:  2016-12-30       Impact factor: 13.837

2.  Molecular cloning of a non-inactivating proton-gated Na+ channel specific for sensory neurons.

Authors:  R Waldmann; F Bassilana; J de Weille; G Champigny; C Heurteaux; M Lazdunski
Journal:  J Biol Chem       Date:  1997-08-22       Impact factor: 5.157

Review 3.  Peptide-gated ion channels and the simple nervous system of Hydra.

Authors:  Stefan Gründer; Marc Assmann
Journal:  J Exp Biol       Date:  2015-02-15       Impact factor: 3.312

4.  Optimization protein productivity of human interleukin-2 through codon usage, gene copy number and intracellular tRNA concentration in CHO cells.

Authors:  Kua-Chun Ou; Chih-Yang Wang; Kuan-Ting Liu; Yi-Ling Chen; Yi-Chen Chen; Ming-Derg Lai; Meng-Chi Yen
Journal:  Biochem Biophys Res Commun       Date:  2014-10-24       Impact factor: 3.575

5.  Sequence, structure, and codon preference of the Drosophila ribosomal protein 49 gene.

Authors:  P O O'Connell; M Rosbash
Journal:  Nucleic Acids Res       Date:  1984-07-11       Impact factor: 16.971

Review 6.  Codon usage and tRNA content in unicellular and multicellular organisms.

Authors:  T Ikemura
Journal:  Mol Biol Evol       Date:  1985-01       Impact factor: 16.240

7.  Acid-sensing ion channel 3 matches the acid-gated current in cardiac ischemia-sensing neurons.

Authors:  S P Sutherland; C J Benson; J P Adelman; E W McCleskey
Journal:  Proc Natl Acad Sci U S A       Date:  2000-12-19       Impact factor: 11.205

8.  Codon preference optimization increases prokaryotic cystatin C expression.

Authors:  Qing Wang; Cui Mei; Honghua Zhen; Jess Zhu
Journal:  J Biomed Biotechnol       Date:  2012-10-03

9.  Convergent evolution of sodium ion selectivity in metazoan neuronal signaling.

Authors:  Maya Gur Barzilai; Adam M Reitzel; Johanna E M Kraus; Dalia Gordon; Ulrich Technau; Michael Gurevitz; Yehu Moran
Journal:  Cell Rep       Date:  2012-07-26       Impact factor: 9.423

10.  Functional Characterization of Cnidarian HCN Channels Points to an Early Evolution of Ih.

Authors:  Emma C Baker; Michael J Layden; Damian B van Rossum; Bishoy Kamel; Monica Medina; Eboni Simpson; Timothy Jegla
Journal:  PLoS One       Date:  2015-11-10       Impact factor: 3.240

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