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The Second Transmembrane Domain of P2X7 Contributes to Dilated Pore Formation

Sun, C and Heid, ME and Keyel, PA and Salter, RD (2013) The Second Transmembrane Domain of P2X7 Contributes to Dilated Pore Formation. PLoS ONE, 8 (4).

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Abstract

Activation of the purinergic receptor P2X7 leads to the cellular permeability of low molecular weight cations. To determine which domains of P2X7 are necessary for this permeability, we exchanged either the C-terminus or portions of the second transmembrane domain (TM2) with those in P2X1 or P2X4. Replacement of the C-terminus of P2X7 with either P2X1 or P2X4 prevented surface expression of the chimeric receptor. Similarly, chimeric P2X7 containing TM2 from P2X1 or P2X4 had reduced surface expression and no permeability to cationic dyes. Exchanging the N-terminal 10 residues or C-terminal 14 residues of the P2X7 TM2 with the corresponding region of P2X1 TM2 partially restored surface expression and limited pore permeability. To further probe TM2 structure, we replaced single residues in P2X7 TM2 with those in P2X1 or P2X4. We identified multiple substitutions that drastically changed pore permeability without altering surface expression. Three substitutions (Q332P, Y336T, and Y343L) individually reduced pore formation as indicated by decreased dye uptake and also reduced membrane blebbing in response to ATP exposure. Three others substitutions, V335T, S342G, and S342A each enhanced dye uptake, membrane blebbing and cell death. Our results demonstrate a critical role for the TM2 domain of P2X7 in receptor function, and provide a structural basis for differences between purinergic receptors. © 2013 Sun et al.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Sun, Cchs79@pitt.eduCHS79
Heid, ME
Keyel, PA
Salter, RD
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorBobé, PierreUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 17 April 2013
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 8
Number: 4
DOI or Unique Handle: 10.1371/journal.pone.0061886
Schools and Programs: School of Medicine > Immunology
Refereed: Yes
Date Deposited: 14 May 2013 16:10
Last Modified: 29 Jan 2019 15:55
URI: http://d-scholarship.pitt.edu/id/eprint/18640

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