TY - JOUR
T1 - Atp and large signaling metabolites flux through caspase-activated pannexin 1 channels
AU - Narahari, Adishesh K.
AU - Kreutzberger, Alex J.B.
AU - Gaete, Pablo S.
AU - Chiu, Yu Hsin
AU - Leonhardt, Susan A.
AU - Medina, Christopher B.
AU - Jin, Xueyao
AU - Oleniacz, Patrycja W.
AU - Kiessling, Volker
AU - Barrett, Paula Q.
AU - Ravichandran, Kodi S.
AU - Yeager, Mark
AU - Contreras, Jorge E.
AU - Tamm, Lukas K.
AU - Bayliss, Douglas A.
N1 - Funding Information:
This work was supported by P01 HL120840 (KSR, DAB, MY); R01 HL138241 (PQB); R01 GM099490 (JEC); R01 GM138532 (MY); and R01 HL48908 (MY); and P01 GM072694 (LKT) and R01 GM051329 (LKT). AKN was supported by F30 CA236370, T32 GM007267, and the University of Virginia Whit-field-Randolph Scholarship, and AKN and CBM were supported by T32 GM007055. YHC was supported by MOST 108–2320-B-007–007-MY2. The authors are grateful for sample preparation by Sandra Poulos, liposome preparation advice from Raghavendar Reddy Sanganna Gari and Patrick Seelheim, helpful early advice on liposome bulk dye uptake assays from Joseph A Mindell (NINDS), and the University of Virginia flow cytometry core facility. The authors are grateful to Elizabeth Gonye, Keyong Li, and Yingtang Shi (Bayliss Laboratory) for helpful discussions and suggestions.
Funding Information:
This work was supported by P01 HL120840 (KSR, DAB, MY); R01 HL138241 (PQB); R01 GM099490 (JEC); R01 GM138532 (MY); and R01 HL48908 (MY); and P01 GM072694 (LKT) and R01 GM051329 (LKT). AKN was supported by F30 CA236370, T32 GM007267, and the University of Virginia Whitfield-Randolph Scholarship, and AKN and CBM were supported by T32 GM007055. YHC was supported by MOST 108–2320-B-007–007-MY2. The authors are grateful for sample preparation by Sandra Poulos, liposome preparation advice from Raghavendar Reddy Sanganna Gari and Patrick Seelheim, helpful early advice on liposome bulk dye uptake assays from Joseph A Mindell (NINDS), and the University of Virginia flow cytometry core facility. The authors are grateful to Elizabeth Gonye, Keyong Li, and Yingtang Shi (Bayliss Laboratory) for helpful discussions and suggestions.
Publisher Copyright:
© Narahari et al.
PY - 2021/1
Y1 - 2021/1
N2 - Pannexin 1 (Panx1) is a membrane channel implicated in numerous physiological and pathophysiological processes via its ability to support release of ATP and other cellular metabolites for local intercellular signaling. However, to date, there has been no direct demonstration of large molecule permeation via the Panx1 channel itself, and thus the permselectivity of Panx1 for different molecules remains unknown. To address this, we expressed, purified, and reconstituted Panx1 into proteoliposomes and demonstrated that channel activation by caspase cleavage yields a dye-permeable pore that favors flux of anionic, large-molecule permeants (up to ~1 kDa). Large cationic molecules can also permeate the channel, albeit at a much lower rate. We further show that Panx1 channels provide a molecular pathway for flux of ATP and other anionic (glutamate) and cationic signaling metabolites (spermidine). These results verify large molecule permeation directly through caspase-activated Panx1 channels that can support their many physiological roles.
AB - Pannexin 1 (Panx1) is a membrane channel implicated in numerous physiological and pathophysiological processes via its ability to support release of ATP and other cellular metabolites for local intercellular signaling. However, to date, there has been no direct demonstration of large molecule permeation via the Panx1 channel itself, and thus the permselectivity of Panx1 for different molecules remains unknown. To address this, we expressed, purified, and reconstituted Panx1 into proteoliposomes and demonstrated that channel activation by caspase cleavage yields a dye-permeable pore that favors flux of anionic, large-molecule permeants (up to ~1 kDa). Large cationic molecules can also permeate the channel, albeit at a much lower rate. We further show that Panx1 channels provide a molecular pathway for flux of ATP and other anionic (glutamate) and cationic signaling metabolites (spermidine). These results verify large molecule permeation directly through caspase-activated Panx1 channels that can support their many physiological roles.
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U2 - 10.7554/ELIFE.64787
DO - 10.7554/ELIFE.64787
M3 - Article
C2 - 33410749
AN - SCOPUS:85100005487
SN - 2050-084X
VL - 10
SP - 1
EP - 21
JO - eLife
JF - eLife
M1 - e64787
ER -