285
16. Seal plates with parafilm to protect cell culture from contamination and drying. Orientation mark will help to identify the
original colony on the mother plate.
17. Effective irradiation to allow yeast functional complementation of BLINK light-driven potassium channel [5]:
210 ± 20 μW/cm
2
using Royal Blue Light (λ = 447 nm).
18. The average yield of differentially growing colonies obtained is
up to 20 per plate.
Acknowledgments
This work was supported by MAE (Ministero Affari Esteri) grant
n. 01467532013-06-27 and ERC 2015 AdG n. 695078 noMAGIC
to A.M.
References
1. Ohndorf UM, MacKinnon R (2005)
Construction of a cyclic nucleotide-gated KcsA
K+ channel. J Mol Biol 350:857–865
2. McCoy JG, Rusinova R, Kim DM, Kowal J,
Banerjee S, Jaramillo Cartagena A, Thompson
AN, Kolmakova-Partensky L, Stahlberg H,
Andersen OS, Nimigean CM (2014) A KcsA/
MloK1 chimeric ion channel has lipiddependent ligand-binding energetics. J Biol
Chem 289(14):9535–9546
3. Arrigoni C, Schroeder I, Romani G, Van Etten
JL, Thiel G, Moroni A (2013) The voltagesensing domain of a phosphatase gates the pore
of a potassium channel. J Gen Physiol
141:389–395
4. Cosentino C, Alberio L, Gazzarrini S, Aquila
M, Romano E, Cermenati S, Zuccolini P et al
(2015) Engineering of a light-gated potassium
channel. Science 348:707–710
5. Christie JM, Arvai AS, Baxtger KJ, Heilmann
M, Pratt AJ, O’Hara A, Kelly SM et al (2012)
Plant UVR8 photoreceptor senses UV-B by
tryptophan-mediated disruption of cross-dimer
salt bridges. Science 335:1492–1496
6. Minor DL, Masseling SJ, Jan YN, Jan LY (1999)
Transmembrane structure of an inwardly rectifying potassium channel. Cell 96:879–891
7. Chatelain FC, Gazzarrini S, Fujiwara Y,
Arrigoni C, Domigan C, Ferrara G, Pantoja C,
Thiel G, Moroni A, Minor DL Jr (2009)
Selection of inhibitor-resistant viral potassium
channels identifies a selectivity filter site that
affects barium and amantadine block. PLoS
One 4(10):e7496
8. Tang W, Ruknudin A, Yang WP, Shaw SY,
Knickerbocker A, Kurtz S (1995) Functional
expression of a vertebrate inwardly rectifying K
+
channel in yeast. Mol Biol Cell 6:1231–1240
Engineering Light-Regulated K
+
Channels
16. Seal plates with parafilm to protect cell culture from contamination and drying. Orientation mark will help to identify the
original colony on the mother plate.
17. Effective irradiation to allow yeast functional complementation of BLINK light-driven potassium channel [5]:
210 ± 20 μW/cm
2
using Royal Blue Light (λ = 447 nm).
18. The average yield of differentially growing colonies obtained is
up to 20 per plate.
Acknowledgments
This work was supported by MAE (Ministero Affari Esteri) grant
n. 01467532013-06-27 and ERC 2015 AdG n. 695078 noMAGIC
to A.M.
References
1. Ohndorf UM, MacKinnon R (2005)
Construction of a cyclic nucleotide-gated KcsA
K+ channel. J Mol Biol 350:857–865
2. McCoy JG, Rusinova R, Kim DM, Kowal J,
Banerjee S, Jaramillo Cartagena A, Thompson
AN, Kolmakova-Partensky L, Stahlberg H,
Andersen OS, Nimigean CM (2014) A KcsA/
MloK1 chimeric ion channel has lipiddependent ligand-binding energetics. J Biol
Chem 289(14):9535–9546
3. Arrigoni C, Schroeder I, Romani G, Van Etten
JL, Thiel G, Moroni A (2013) The voltagesensing domain of a phosphatase gates the pore
of a potassium channel. J Gen Physiol
141:389–395
4. Cosentino C, Alberio L, Gazzarrini S, Aquila
M, Romano E, Cermenati S, Zuccolini P et al
(2015) Engineering of a light-gated potassium
channel. Science 348:707–710
5. Christie JM, Arvai AS, Baxtger KJ, Heilmann
M, Pratt AJ, O’Hara A, Kelly SM et al (2012)
Plant UVR8 photoreceptor senses UV-B by
tryptophan-mediated disruption of cross-dimer
salt bridges. Science 335:1492–1496
6. Minor DL, Masseling SJ, Jan YN, Jan LY (1999)
Transmembrane structure of an inwardly rectifying potassium channel. Cell 96:879–891
7. Chatelain FC, Gazzarrini S, Fujiwara Y,
Arrigoni C, Domigan C, Ferrara G, Pantoja C,
Thiel G, Moroni A, Minor DL Jr (2009)
Selection of inhibitor-resistant viral potassium
channels identifies a selectivity filter site that
affects barium and amantadine block. PLoS
One 4(10):e7496
8. Tang W, Ruknudin A, Yang WP, Shaw SY,
Knickerbocker A, Kurtz S (1995) Functional
expression of a vertebrate inwardly rectifying K
+
channel in yeast. Mol Biol Cell 6:1231–1240
Engineering Light-Regulated K
+
Channels