IndraLab
Statements
sparser
"Thermodynamic measurements of ion binding using isothermal titration calorimetry (ITC) report a K d of ∼0.43 mM for K + binding. xref Electrophysiological experiments on the KcsA-Kv1.3 chimera report two high affinity binding sites in the pore: an inner site that has a K d of 6.5 µM and the outer site has a K d of 0.9 mM."
sparser
"Comparison of properties of GFP-L2-AgTx2 and its structural precursor His6-GFP-L2-AgTx2 shows that any of these ligands can be efficiently used as a selective fluorescent probe in the bioengineering analytical system on the basis of KcsA-Kv1.3 channel to search for Kv1.3 channel blockers (both among individual compounds and in complex mixtures) and to estimate their activity ( xref E,F, xref )."
sparser
"Rather high concentration of HTX or MTX was required to displace agitoxin 2 from the KcsA-Kv1.3 channel because affinity of agitoxin 2 to the Kv1.3 binding site was much higher xref than that of the tested peptides in accordance with the properties of wild type HTX xref and MTX. xref "
sparser
"To examine the capability of the Nano-oscillator platform for measuring small molecule binding kinetics and demonstrate its potential application in drug screening, we studied the binding of a small molecule drug candidate 1 (4-(2-ethylpiperidin-1-yl)-2-methyl-6-phenyl-5H-pyrrolo[3,2-d] pyrimidine, MW = 320 Da) to a chimeric ion channel protein KcsA-Kv1.3 ( xref ), which plays an important role in autoimmune diseases. xref To stabilize the membrane protein in aqueous solution, KcsA-Kv1.3 was encapsulated in a Nanodisc consisting of a nano-scale lipid bilayer disc fastened by a membrane scaffold protein."
sparser
"To characterize potential of GFP-L2-AgTx2 and His6-GFP-L2-AgTx2 as a component of the KcsA-Kv1.3-based bioengineering system for searching Kv1.3 channel ligands in complex mixtures, the response of complexes between KcsA-Kv1.3 and GFP-L2-AgTx2 or His6-GFP-L2-AgTx2 to addition of scorpion venoms was measured ( xref D)."
sparser
"We also measured a peptide blocker of KcsA-Kv1.3, ShK, which is more potent than 1 and has been considered as a model by pharmaceutical companies for developing immunosuppressant drugs. xref The binding curves and fittings are shown in xref , and k a , k d , and K D were determined to be 2.0×10 7 M −1 s −1 , 4.0×10 −3 s −1 , and 0.20 nM, respectively."
sparser
"To verify the specific binding of ShK, two additional experiments were carried out by adding ShK to empty Nanodisc and adding IgG to KcsA-Kv1.3 Nanodisc, and neither of them produced detectable oscillation amplitude changes ( xref , curves marked with control 1 and control 2, respectively)."
sparser
"To estimate applicability of GFP–MgTx for recognition of Kv1.3 blockers, we studied displacement of GFP–MgTx from the complexes with KcsA–Kv1.3 by known pore blockers of Kv1.3 channel, namely, two peptide toxins, ChTx and HgTx1, and a low-molecular-weight blocker tetraethylammonium (TEA), which has millimolar affinity to Kv1.3 [ xref ] ( xref )."
sparser
"HTX is a new Kv1.3 channel blocker from the scorpion Heterometrus laoticus . xref MTX is a high affinity blocker of Kv1.2 and K Ca channels xref as well as a moderate affinity blocker of Kv1.3 channel. xref To confirm correct folding of recombinant HTX and MTX, we have tested their ability to bind to the Kv1.3-channel binding site formed in the hybrid KcsA-Kv1.3 channel."
sparser
"To do this, a bioengineering system was used, which was based on a KcsA-Kv1.3 channel embedded in the membrane of spheroplasts and fluorescently labeled peptide blocker agitoxin 2. xref This system is a reliable analytical tool to search for Kv1.3 blockers in complex mixtures and among individual compounds as well as to characterize their activities. xref Using this system as described elsewhere, xref we have observed displacement of fluorescent agitoxin 2 from the Kv1.3-channel binding site by HTX and MTX ( xref )."