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AM7A protein product. Ric-3 was detected in SH-EP1– h7-Ric-3 cell samples and met all inclusion criteria, but it was associated with a borderline probability score of 88. This may reflect the fact that Ric-3 is only transiently associated with 7-nAChRs. Not all 3PO 7-nAChRs will be interacting with Ric-3 at the time of -bgtx-affinity bead isolation. The 2MNaCl washes for all samples were also analyzed to confirm that 7-nAChRs were not eluted during the washing of material bound to -bgtx-affinity beads. Neither 7-nAChR peptides nor Ric-3 peptides were identified in the 2MNaCl bead wash from SH-EP1-h7-Ric-3 cell samples. Identification of the 7-nAChR in SH-EP1-h7-Ric-3 and SH-EP1-h7 cell samples confirms that 7-nAChRs were eluted from the -bgtx-affinity beads by the cholinergic agonist carbachol. Neither 7-nAChR nor Ric-3 peptides were identified in carbachol-eluted samples prepared from SH-EP1 cells, which lack expression of both proteins. A peptide corresponding to 7-nAChR subunits was identified in -bgtx-enriched samples of both SH-EP1-h7-Ric-3 and SH-EP1-h7 cell lines. Data analysis was performed using ProteoIQ version 2.7 Protein inclusion criteria include 1 protein FDR, minimum peptide length of six amino acids 90 probability, identification in 2 or more of 5 replicates , and 0 probability in controls. FDRs were determined using the PROVALT 219832-49-2 algorithm and probabilities were determined with the ProteinProphet algorithm through ProteoIQ analysis. Only Top and Co-Top identifications were considered. Proteins identified in our analysis of the 7-nAChR interactome are most likely components of large protein complexes and may either be associating directly with the receptor or with another member of the complex. Comparison of carbachol-eluted proteins from SH-EP1-h7-Ric-3 and SH-EP1-h7 identified thirty-nine Ric-3-promoted 7-nAChR associated proteins. Fourteen of the thirty-nine proteins identified as Ric-3-mediated have previously been reported as associated with a cellular process known to affect protein expression. These fourteen Ric-3-mediated associated proteins may be directly or indirectly recruited by Ric-3 to facilitate receptor assembly and targeting. In addi

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Author: calcimimeticagent