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. 2023 May 26;12:e84645. doi: 10.7554/eLife.84645

Figure 2. The intracellular juxtamembrane (ICJM) region of the prolactin receptor (PRLR) interacts with PI(4,5)P2.

(A) Overview of investigated PRLR variants. (B) Secondary chemical shifts (SCSs) of transmembrane domain (TMD)-intracellular domain (ICD)F206-S270 reconstituted in 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) micelles. (C) Correlation plot of the SCSs of ICDG236-Q396 plotted against those of TMD-ICDF206-S270. (D) 15N,1H-HSQC spectra of 15N-ICDK235-G313 titrated with 5×, 10×, and 25× molar excess of C8-PI(4,5)P2. (E) Structure of C8-PI(4,5)P2. (F) Backbone amide chemical shift perturbations (CSPs) and peak intensity changes upon addition of C8-PI(4,5)P2 to 15N-ICDK235-G313 plotted against residue number. (G) Top: Far-UV CD spectra of Pep1 titrated with C8-PI(4,5)P2 or in 65% trifluorethanol (TFE). Middle: Far-UV CD spectra of Pep1 in the presence of 5 x-38x C8-PI(4,5)P2 subtracted with the spectrum of Pep1 in the absence of C8-PI(4,5)P2. Bottom: Far-UV CD spectra of Pep2 titrated with C8-PI(4,5)P2 or in 65% TFE.

Figure 2.

Figure 2—figure supplement 1. 15N, 1H-HSQC spectra.

Figure 2—figure supplement 1.

(A) transmembrane domain (TMD)F206-V240 and TMD-intracellular domain (ICD)F206-S270 in 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) micelles, and (B) TMD-ICDF206-S270 in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) small unilamellar vesicles (SUVs).
Figure 2—figure supplement 2. Cα secondary chemical shifts of intracellular domain (ICD)G236-Q396.

Figure 2—figure supplement 2.