Generation of functional ion-channel tools by E3 targeting.
Here we describe a strategy for generating ion-channel inhibitors. It takes advantage of antibody specificity combined with a pattern recognition approach that targets the third extracellular region (E3) of a channel. To test the concept, we first focused on TRPC5, a member of the transient receptor potential (TRP) calcium channel family, the study of which has been hindered by poor pharmacological tools. Extracellular application of E3-targeted anti-TRPC5 antibody led to a specific TRPC5 inhibitor, enabling TRPC5 to be distinguished from its closest family members, and TRPC5 function to be explored in a relatively intractable physiological system. E3 targeting was further applied to voltage-gated sodium channels, leading to discovery of a subtype-specific inhibitor of Na(V)1.5. These examples illustrate the potential power of E3 targeting as a systematic method for producing gene-type specific ion-channel inhibitors for use in routine assays on cells or tissues from a range of species and having therapeutic potential.
Xu
Shang-Zhong
SZ
Membrane and Systems Biology Research Institute, University of Leeds, Leeds, LS2 9JT, UK.
Zeng
Fanning
F
Lei
Ming
M
Li
Jing
J
Gao
Bin
B
Xiong
Chenliang
C
Sivaprasadarao
Asipu
A
Beech
David J
DJ
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D001665:Binding Sites; D016503:Drug Delivery Systems / Q000379:methods*; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology*; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D050052:TRPC Cation Channels / Q000187:drug effects* / Q000502:physiology*
D000911:Antibodies, Monoclonal; D007473:Ion Channels; D050052:TRPC Cation Channels; C491780:TRPC5 protein, human