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A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor : computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and in situ affinity constant in cells
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- Szalai, Alan M. (författare)
- Consejo Nacl Invest Cient & Tecn, Ctr Invest Bionanociencias Elizabeth Jares Erijma, Godoy Cruz 2390, RA-1425 Buenos Aires, DF, Argentina.;Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Quim Organ Analit & Quim, Pabellon 2 Ciudad Univ, RA-1428 Buenos Aires, DF, Argentina.
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- Armando, Natalia G. (författare)
- Consejo Nacl Invest Cient & Tecn, Inst Invest Biomed Buenos Aires IBioBA, Max Planck Soc, Partner Inst, Godoy Cruz 2390, RA-1425 Buenos Aires, DF, Argentina.;Univ Nacl Quilmes, Dept Ciencias & Tecnol, Roque Saenz Pena 352, RA-1876 Bernal, Buenos Aires, Argentina.
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- Barabas, Federico M. (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Consejo Nacl Invest Cient & Tecn, Ctr Invest Bionanociencias Elizabeth Jares Erijma, Godoy Cruz 2390, RA-1425 Buenos Aires, DF, Argentina.;Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Fis, Pabellon 1 Ciudad Univ, RA-1428 Buenos Aires, DF, Argentina
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- Consejo Nacl Invest Cient & Tecn, Ctr Invest Bionanociencias Elizabeth Jares Erijma, Godoy Cruz 2390, RA-1425 Buenos Aires, DF, Argentina;Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Quim Organ Analit & Quim, Pabellon 2 Ciudad Univ, RA-1428 Buenos Aires, DF, Argentina. Consejo Nacl Invest Cient & Tecn, Inst Invest Biomed Buenos Aires IBioBA, Max Planck Soc, Partner Inst, Godoy Cruz 2390, RA-1425 Buenos Aires, DF, Argentina.;Univ Nacl Quilmes, Dept Ciencias & Tecnol, Roque Saenz Pena 352, RA-1876 Bernal, Buenos Aires, Argentina. (creator_code:org_t)
- 2018
- 2018
- Engelska.
- Ingår i: Physical Chemistry, Chemical Physics - PCCP. - : Royal Society of Chemistry. - 1463-9076 .- 1463-9084. ; 10:46, s. 29212-29220
- Tidskriftsartikel (refereegranskat)
Abstract
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- Class B G protein-coupled receptors (GPCRs) are involved in a variety of human pathophysiological states. These groups of membrane receptors are less studied than class A GPCRs due to the lack of structural information, delayed small molecule drug discovery, and scarce fluorescence detection tools available. The class B corticotropin-releasing hormone type 1 receptor (CRHR1) is a key player in the stress response whose dysregulation is critically involved in stress-related disorders: psychiatric conditions (i.e. depression, anxiety, and addictions), neuroendocrinological alterations, and neurodegenerative diseases. Here, we present a strategy to label GPCRs with a small fluorescent antagonist that permits the observation of the receptor in live cells through stochastic optical reconstruction microscopy (STORM) with 23 nm resolution. The marker, an aza-BODIPY derivative, was designed based on computational docking studies, then synthesized, and finally tested in biological cells. Experiments on hippocampal neurons demonstrate antagonist effects in similar concentrations as the well-established antagonist CP-376395. A quantitative analysis of two color STORM images enabled the determination of the binding affinity of the new marker in the cellular environment.
Ämnesord
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
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