Analysis of plasma membrane integrity by fluorescent detection of Tl(+) uptake.
AuthorsBowman AM, Nesin OM, Pakhomova ON, Pakhomov AG,
JournalJ Membr Biol
PubMed ID20623351
'The exclusion of polar dyes by healthy cells is widely employed as a simple and reliable test for cell membrane integrity. However, commonly used dyes (propidium, Yo-Pro-1, trypan blue) cannot detect membrane defects which are smaller than the dye molecule itself, such as nanopores that form by exposure to ultrashort ... More
The Ste20 kinases Ste20-related proline-alanine-rich kinase and oxidative-stress response 1 regulate NKCC1 function in sensory neurons.
AuthorsGeng Y, Hoke A, Delpire E,
JournalJ Biol Chem
PubMed ID19307180
'NKCC1 is highly expressed in dorsal root ganglion neurons, where it is involved in gating sensory information. In a recent study, it was shown that peripheral nerve injury results in increased NKCC1 activity, not due to an increase in cotransporter expression, but to increased phosphorylation of the cotransporter (Pieraut, S., ... More
Discovery of novel allosteric modulators of metabotropic glutamate receptor subtype 5 reveals chemical and functional diversity and in vivo activity in rat behavioral models of anxiolytic and antipsychotic activity.
AuthorsRodriguez AL, Grier MD, Jones CK, Herman EJ, Kane AS, Smith RL, Williams R, Zhou Y, Marlo JE, Days EL, Blatt TN, Jadhav S, Menon UN, Vinson PN, Rook JM, Stauffer SR, Niswender CM, Lindsley CW, Weaver CD, Conn PJ,
JournalMol Pharmacol
PubMed ID20923853
'Modulators of metabotropic glutamate receptor subtype 5 (mGluR5) may provide novel treatments for multiple central nervous system (CNS) disorders, including anxiety and schizophrenia. Although compounds have been developed to better understand the physiological roles of mGluR5 and potential usefulness for the treatment of these disorders, there are limitations in the ... More
Development of a selective small-molecule inhibitor of Kir1.1, the renal outer medullary potassium channel.
'The renal outer medullary potassium (K+) channel, ROMK (Kir1.1), is a putative drug target for a novel class of loop diuretic that would lower blood volume and pressure without causing hypokalemia. However, the lack of selective ROMK inhibitors has hindered efforts to assess its therapeutic potential. In a high-throughput screen ... More
Lipid nanopores can form a stable, ion channel-like conduction pathway in cell membrane.
AuthorsPakhomov AG, Bowman AM, Ibey BL, Andre FM, Pakhomova ON, Schoenbach KH,
JournalBiochem Biophys Res Commun
PubMed ID19450553
Cell permeabilization by electric pulses (EPs), or electroporation, has been well established as a tool to indiscriminately increase membrane flows of water solutes down the concentration and voltage gradients. However, we found that EPs of nanosecond duration (nsEPs) trigger formation of voltage-sensitive and inward-rectifying membrane pores. NsEP-treated cells remain mostly ... More
The K(+) channel GIRK2 is both necessary and sufficient for peripheral opioid-mediated analgesia.
AuthorsNockemann D, Rouault M, Labuz D, Hublitz P, McKnelly K, Reis FC, Stein C, Heppenstall PA,
Journal
PubMed ID23818182
The use of opioid agonists acting outside the central nervous system (CNS) is a promising therapeutic strategy for pain control that avoids deleterious central side effects such as apnea and addiction. In human clinical trials and rat models of inflammatory pain, peripherally restricted opioids have repeatedly shown powerful analgesic effects; ... More
Lipids as tumoricidal components of human a-lactalbumin made lethal to tumor cells (HAMLET): unique and shared effects on signaling and death.
AuthorsHo JC, Storm P, Rydström A, Bowen B, Alsin F, Sullivan L, Ambite I, Mok KH, Northen T, Svanborg C,
JournalJ Biol Chem
PubMed ID23629662
Long-chain fatty acids are internalized by receptor-mediated mechanisms or receptor-independent diffusion across cytoplasmic membranes and are utilized as nutrients, building blocks, and signaling intermediates. Here we describe how the association of long-chain fatty acids to a partially unfolded, extracellular protein can alter the presentation to target cells and cellular effects. ... More
hERG K(+) channels: structure, function, and clinical significance.
AuthorsVandenberg JI, Perry MD, Perrin MJ, Mann SA, Ke Y, Hill AP,
JournalPhysiol Rev
PubMed ID22988594
The human ether-a-go-go related gene (hERG) encodes the pore-forming subunit of the rapid component of the delayed rectifier K(+) channel, Kv11.1, which are expressed in the heart, various brain regions, smooth muscle cells, endocrine cells, and a wide range of tumor cell lines. However, it is the role that Kv11.1 ... More
A novel mitochondrial K(ATP) channel assay.
AuthorsWojtovich AP, Williams DM, Karcz MK, Lopes CM, Gray DA, Nehrke KW, Brookes PS,
JournalCirc Res
PubMed ID20185796
The mitochondrial ATP sensitive potassium channel (mK(ATP)) is implicated in cardioprotection by ischemic preconditioning (IPC), but the molecular identity of the channel remains controversial. The validity of current methods to assay mK(ATP) activity is disputed. We sought to develop novel methods to assay mK(ATP) activity and its regulation. Using a ... More
An ion channel library for drug discovery and safety screening on automated platforms.
AuthorsWible BA, Kuryshev YA, Smith SS, Liu Z, Brown AM,
JournalAssay Drug Dev Technol
PubMed ID19090691
Ion channels represent the third largest class of targets in drug discovery after G-protein coupled receptors and kinases. In spite of this ranking, ion channels continue to be under exploited as drug targets compared with the other two groups for several reasons. First, with 400 ion channel genes and an ... More
A new homogeneous high-throughput screening assay for profiling compound activity on the human ether-a-go-go-related gene channel.
AuthorsTitus SA, Beacham D, Shahane SA, Southall N, Xia M, Huang R, Hooten E, Zhao Y, Shou L, Austin CP, Zheng W,
JournalAnal Biochem
PubMed ID19583963
Long QT syndrome, either inherited or acquired from drug treatments, can result in ventricular arrhythmia (torsade de pointes) and sudden death. Human ether-a-go-go-related gene (hERG) channel inhibition by drugs is now recognized as a common reason for the acquired form of long QT syndrome. It has been reported that more ... More
Cell-based potassium ion channel screening using the FluxOR assay.
AuthorsBeacham DW, Blackmer T, O' Grady M, Hanson GT,
JournalJ Biomol Screen
PubMed ID20208034
FluxOR technology is a cell-based assay used for high-throughput screening measurements of potassium channel activity. Using thallium influx as a surrogate indicator of potassium ion channel activity, the FluxOR Potassium Ion Channel Assay is based on the activation of a novel fluorescent dye. This indicator reports channel activity with a ... More
A thallium-sensitive, fluorescence-based assay for detecting and characterizing potassium channel modulators in mammalian cells.
Potassium channels have been identified as targets for a large number of therapeutic indications. The ability to use a high-throughput functional assay for the detection and characterization of small-molecule modulators of potassium channels is very desirable. However, present techniques capable of screening very large chemical libraries are limited in terms ... More