bioimagesUnknownTheresia GutmannThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201711047JCB

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1T, T-shaped IR in one nanodisc; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 10,619 particles. The class averages were visually assigned to show a T-shaped IR in one nanodisc (1T), an L-shaped monomeric IR (L), or none of these (X). (C) The 202 averages resulting from 5 ISAC generations. The averages shown in Fig. 3 D are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 2U, U-shaped IR in two nanodiscs; 1T, T-shaped IR in one nanodisc; 2T, T-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 13,304 particles. The class averages were visually assigned to show a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), a T-shaped IR in two nanodiscs (2T), an L-shaped monomeric IR (L), or none of these (X). (C) The 286 averages resulting from 13 ISAC generations. The averages shown in Fig. S4 B are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1U, U-shaped IR in one nanodisc; 2U, U-shaped IR in two nanodiscs; 1T, T-shaped IR in one nanodisc; 2T, T-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 10,858 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), a T-shaped IR in two nanodiscs (2T), an L-shaped monomeric IR (L), or none of these (X). (C) The 230 averages resulting from 7 ISAC generations. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1U, U-shaped IR in one nanodisc; 1T, T-shaped IR in one nanodisc; II: II-shaped IR in one nanodisc; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 14,820 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a T-shaped IR in one nanodisc (1T), an II-shaped IR in one nanodisc (II), an L-shaped monomeric IR (L), or none of these (X). (C) The 307 averages resulting from 9 ISAC generations. The averages shown in Fig. 3 C are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1T, T-shaped IR in one nanodisc; 2U, U-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 5503 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), an L-shaped monomeric IR (L), or none of these (X). (C) The 112 averages resulting from 6 ISAC generations. The averages shown in Fig. S4 D are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 2U, U-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 11,493 particles. The class averages were visually assigned to show a U-shaped IR in two nanodiscs (2U), an L-shaped monomeric IR (L), or none of these (X). (C) The 241 averages resulting from 9 ISAC generations. The averages shown in Fig. S4 A are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1T, T-shaped IR in one nanodisc; 2T, T-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 10,239 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), a T-shaped IR in two nanodiscs (2T), an L-shaped monomeric IR (L), or none of these (X). (C) The 166 averages resulting from 3 ISAC generations. The averages shown in Fig. S74C are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1U, U-shaped IR in one nanodisc; 2U, U-shaped IR in two nanodiscs; 1T, T-shaped IR in one nanodisc; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 10,634 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), an L-shaped monomeric IR (L), or none of these (X). (C) The 215 averages resulting from 7 ISAC generations. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 2U, U-shaped IR in two nanodiscs; 1T, T-shaped IR in one nanodisc; 2T: T-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 14,041 particles. The class averages were visually assigned to show a U-shaped IR in two nanodiscs (2U), a T-shaped IR in one nanodisc (1T), a T-shaped IR in two nanodiscs (2T), an L-shaped monomeric IR (L), or none of these (X). (C) The 299 averages resulting from 9 ISAC generations. The averages shown in Fig. 3 B are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

(A) Selected area of a raw negative-stain EM image. Some representative particles are circled: 1U, U-shaped IR in one nanodisc; 2U, U-shaped IR in two nanodiscs; L, monomeric IR in one nanodisc. (B) The 200 class averages obtained from K-means classification of 13,800 particles. The class averages were visually assigned to show a U-shaped IR in one nanodisc (1U), a U-shaped IR in two nanodiscs (2U), an L-shaped IR (L), or none of these (X). (C) The 297 averages resulting from 8 ISAC generations. The averages shown in Fig. 3 A are marked with an asterisk. Side length of individual averages in panels B and C: 47.7 nm.

bioimagesD6, Orig. Data to Fig 3ED10, Orig. Data to Fig S4DImage 643142 (D1, Orig. Data to Fig 3A - None)D9, Orig. Data to Fig S4CD2, Orig. Data to Fig 3B and 3ED5, Orig. Data to Fig 3E - NoneImage 643147 (D6, Orig. Data to Fig 3E - None)D8, Orig. Data to Fig S4B - NoneD1, Orig. Data to Fig 3AD3, Orig. Data to Fig 3CD4, Orig. Data to Fig 3D - NoneD7, Orig. Data to Fig S4AImage 643143 (D2, Orig. Data to Fig 3B and 3E - None)D9, Orig. Data to Fig S4C - NoneImage 643150 (D9, Orig. Data to Fig S4C - None)D4, Orig. Data to Fig 3DImage 643149 (D8, Orig. Data to Fig S4B - None)D7, Orig. Data to Fig S4A - NoneD3, Orig. Data to Fig 3C - NoneD8, Orig. Data to Fig S4BD10, Orig. Data to Fig S4D - NoneD5, Orig. Data to Fig 3EImage 643146 (D5, Orig. Data to Fig 3E - None)Image 643151 (D10, Orig. Data to Fig S4D - None)Image 643144 (D3, Orig. Data to Fig 3C - None)Image 643148 (D7, Orig. Data to Fig S4A - None)D1, Orig. Data to Fig 3A - NoneD2, Orig. Data to Fig 3B and 3E - NoneImage 643145 (D4, Orig. Data to Fig 3D - None)D6, Orig. Data to Fig 3E - NoneMichal GrzybekÜnal CoskunThomas WalzTheresia GutmannKelly H. KimfalseVisualization of ligand-induced transmembrane signaling in the full-length human insulin receptorInsulin receptor (IR) signaling plays a critical role in the regulation of metabolism and growth in multicellular organisms. IRs are unique among receptor tyrosine kinases in that they exist exclusively as covalent (αβ)<jats:sub>2</jats:sub> homodimers at the cell surface. Transmembrane signaling by the IR can therefore not be based on ligand-induced dimerization as such but must involve structural changes within the existing receptor dimer. In this study, using glycosylated full-length human IR reconstituted into lipid nanodiscs, we show by single-particle electron microscopy that insulin binding to the dimeric receptor converts its ectodomain from an inverted U-shaped conformation to a T-shaped conformation. This structural rearrangement of the ectodomain propagates to the transmembrane 2018-02-16T11:29:30Z2018-11-29T11:29:30Z2018-11-29T11:29:30ZS-JCBD-20171104710.1083/jcb.201711047