Találati lista | Tudóstér

001-es BibID:	BIBFORM091018
Első szerző:	Tajti Gábor (gyógyszerész, biofizikus, sejtbiológus)
Cím:	Immunomagnetic separation is a suitable method for electrophysiology and ion channel pharmacology studies on T cells / Gabor Tajti, Tibor Gabor Szanto, Agota Csoti, Greta Racz, César Evaristo, Peter Hajdu, Gyorgy Panyi
Dátum:	2021
ISSN:	1933-6950 1933-6969
Megjegyzések:	Ion channels play pivotal role in the physiological and pathological function of immune cells. As immune cells represent a functionally diverse population, subtype-specific functional studies, such as single-cell electrophysiology require proper subset identification and separation. Magneticactivated cell sorting (MACS) techniques provide an alternative to fluorescence-activated cell sorting (FACS), however, the potential impact of MACS-related beads on the biophysical and pharmacological properties of the ion channels were not studied yet. We studied the aforementioned properties of the voltage-gated Kv1.3 K+ channel in activated CD4+ T-cells as well as the membrane capacitance using whole-cell patch-clamp following immunomagnetic positive separation, using the REAlease? kit. This kit allows three experimental configurations: bead-bound configuration, bead-free configuration following the removal of magnetic beads, and the labelfree configuration following removal of CD4 recognizing antibody fragments. As controls, we used FACS separation as well as immunomagnetic negative selection. The membrane capacitance and of the biophysical parameters of Kv1.3 gating, voltage-dependence of steady-state activation and inactivation kinetics of the current were not affected by the presence of MACS-related compounds on the cell surface. We found subtle differences in the activation kinetics of the Kv1.3 current that could not be explained by the presence of MACS-related compounds. Neither the equilibrium block of Kv1.3 by TEA+ or charybdotoxin (ChTx) nor the kinetics of ChTx block are affected by the presence of the magnetics beads on the cell surface. Based on our results MACS is a suitable method to separate cells for studying ion channels in non-excitable cells, such as T-lymphocytes.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk Kv1.3 cd4+ T-cell magneticactivated cell sorting macs immunomagnetic separation fluorescenceactivated cell sorting facs
Megjelenés:	Channels. - 15 : 1 (2021), p. 53-66. -
További szerzők:	Szántó Gábor Tibor (1980-) (vegyész) Csóti Ágota (1989-) (biológus) Rácz Gréta Evaristo, César Hajdu Péter (1975-) (biofizikus) Panyi György (1966-) (biofizikus)
Pályázati támogatás:	OTKA K119417 Egyéb EFOP-3.6.2-16-2017-00006 EFOP GINOP-2.3.2-15-2016-00015 GINOP NKFIH K128525 Egyéb Bolyai Research Fellowship Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
Borító:
Saját polcon:

001-es BibID:	BIBFORM105306
035-os BibID:	(WoS)000883005300001 (Scopus)85141634111
Első szerző:	Wai, Dorothy C. C.
Cím:	A Fluorescent Peptide Toxin for Selective Visualization of the Voltage-Gated Potassium Channel Kv1.3 / Wai Dorothy C. C., Naseem Muhammad Umair, Mocsár Gábor, Babu Reddiar Sanjeevini, Pan Yijun, Csoti Agota, Hajdu Peter, Nowell Cameron, Nicolazzo Joseph A., Panyi Gyorgy, Norton Raymond S.
Dátum:	2022
ISSN:	1043-1802
Megjegyzések:	Upregulation of the voltage-gated potassium channel K(V)1.3 is implicated in a range of autoimmune and neuroinflammatory diseases, including rheumatoid arthritis, psoriasis, multiple sclerosis, and type I diabetes. Understanding the expression, localization, and trafficking of K(V)1.3 in normal and disease states is key to developing targeted immunomodulatory therapies. HsTX1[R14A], an analogue of a 34-residue peptide toxin from the scorpion Heterometrus spinifer, binds K(V)1.3 with high affinity (IC50 of 45 pM) and selectivity (2000-fold for K(V)1.3 over K(V)1.1). We have synthesized a fluorescent analogue of HsTX1[R14A] by N-terminal conjugation of a Cy5 tag. Electrophysiology assays show that Cy5-HsTX1[R14A] retains activity against K(V)1.3 (IC50 similar to 0.9 nM) and selectivity over a range of other potassium channels (K(V)1.2, K(V)1.4, K(V)1.5, K(V)1.6, K(Ca)1.1 and K(Ca)3.1), as well as selectivity against heteromeric channels assembled from K(V)1.3/K(V)1.5 tandem dimers. Live imaging of CHO cells expressing green fluorescent protein-tagged K(V)1.3 shows co-localization of Cy5-HsTX1[R14A] and K(V)1.3 fluorescence signals at the cell membrane. Moreover, flow cytometry demonstrated that Cy5-HsTX1[R14A] can detect K(V)1.3-expressing CHO cells. Stimulation of mouse microglia by lipopolysaccharide, which enhances membrane expression of K(V)1.3, was associated with increased staining by Cy5-HsTX1[R14A], demonstrating that it can be used to identify K(V)1.3 in disease-relevant models of inflammation. Furthermore, the biodistribution of Cy5-HsTX1[R14A] could be monitored using ex vivo fluorescence imaging of organs in mice dosed subcutaneously with the peptide. These results illustrate the utility of Cy5-HsTX1[R14A] as a tool for visualizing K(V)1.3, with broad applicability in fundamental investigations of K(V)1.3 biology, and the validation of novel disease indications where K(V)1.3 inhibition may be of therapeutic value.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk Fluorescence Imaging probes Peptides and proteins Potassium Selectivity
Megjelenés:	Bioconjugate Chemistry. - 33 : 11 (2022), p. 2197-2212. -
További szerzők:	Naseem, Muhammad Umair (1993-) (biofizikus, molekuláris biológus) Mocsár Gábor (1981-) (biofizikus) Babu Reddiar, Sanjeevini Pan, Yijun Csóti Ágota (1989-) (biológus) Hajdu Péter (1975-) (biofizikus) Nowell, Cameron Nicolazzo, Joseph A. Panyi György (1966-) (biofizikus) Norton, Raymond S.
Pályázati támogatás:	K143071 OTKA K128525 OTKA EFOP-3.6.1-16-2016-00022 EFOP GINOP-2.3.2-15-2016-00015 GINOP Stipendium Hungaricum Scholarship by the Tempus Public Foundation Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
Borító:
Saját polcon: