A functional Kv1.2-hERG chimaeric channel expressed in Pichia pastoris

Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to h...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2014-02, Vol.4 (1), p.4201-4201, Article 4201
Main Authors: Dhillon, Mandeep S., Cockcroft, Christopher J., Munsey, Tim, Smith, Kathrine J., Powell, Andrew J., Carter, Paul, Wrighton, David C., Rong, Hong-lin, Yusaf, Shahnaz P., Sivaprasadarao, Asipu
Format: Article
Language:English
Subjects:
631/443/592/75
631/45/269/1151
631/45/612
631/57/2271
82/80
82/83
9/74
Amino Acid Sequence
Animals
Binding sites
Cloning, Molecular - methods
Drugs
HERG protein
Humanities and Social Sciences
Ion channels
Kv1.2 Potassium Channel - chemistry
Kv1.2 Potassium Channel - physiology
Kv1.2 Potassium Channel - ultrastructure
Molecular Conformation
Molecular Sequence Data
multidisciplinary
Oocytes
Pharmaceutical industry
Pharmacology
Physiology
Pichia - genetics
Pichia - metabolism
Potassium
Potassium channels (voltage-gated)
Proteins
Rats
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Science
Structural engineering
Structure-Activity Relationship
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to help explain the molecular basis for the differences in their biophysical properties and pharmacology. This is due to low-level expression of many members in native or heterologous systems. One exception is rat Kv1.2 which has been overexpressed in Pichia pastoris and crystallised. Here, we tested chimaeras of rat Kv1.2 with the hERG channel for function in Xenopus oocytes and for overexpression in Pichia . Chimaera containing the S1–S6 transmembrane region of HERG showed functional and pharmacological properties similar to hERG and could be overexpressed and purified from Pichia . Our results demonstrate that rat Kv1.2 could serve as a surrogate to express difficult-to-overexpress members of the six-transmembrane segment channel family.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep04201