1. Volume 67, Issue 3, Pages 528-534.e3 (August 2017)
Vps34 Kinase Domain Dynamics Regulate the Autophagic PI 3-Kinase Complex 
Goran Stjepanovic, Sulochanadevi Baskaran, Mary G. Lin, James H. Hurley 
Molecular Cell 
Volume 67, Issue 3, Pages e3 (August 2017)
DOI: /j.molcel
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2. Molecular Cell 2017 67, 528-534.e3DOI: (10.1016/j.molcel.2017.07.003)
Copyright © 2017 Elsevier Inc. Terms and Conditions

3. Figure 1 EM Analysis of the Conformational Dynamics
(A) Three-dimensional reconstruction of the PI3KC3-C1 complex with the docked structures shown in a ribbon representation.
(B) Representative class averages of the PI3KC3-C1 complex with the VPS34 HELCAT in range of different conformation states.
(C) Percentage of well-resolved particles sorted into 2D class averages with a classical, V-shaped, WD, paddle, and dislodged VPS34 HELCAT. Schematic and representative class averages of the PI3KC3-C1 complex showing the conformations of the VPS34 HELCAT.
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4. Figure 2 CX-MS Analysis of the Conformational Dynamics
Pi3KC3-C1 was crosslinked with a lysine-specific bifunctional crosslinker, then fragmented by proteolysis, and crosslinked peptides were identified by mass spectrometry.
(A) Inter-subunit crosslinks between subunits of the PI3KC3-C1 complex.
(B) Overview of crosslinks observed within the PI3KC3-C1 complex corresponding to the model generated based on crystal structure of yeast complex I (PDB: 5DFZ). The crosslinks are presented in red (>38 Å) and blue (<38 Å).
See also Table S1 and Movie S1.
Molecular Cell  , e3DOI: ( /j.molcel )
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5. Figure 3 Leashing of the VPS34 HELCAT to VPS15
(A) Schematic of the PI3KC3-C1 complex showing leashed and unleashed construct of the VPS34 HELCAT.
(B) Percentage of well-resolved particles sorted into 2D class averages with a classical, V-shaped, WD, paddle, and dislodged VPS34 HELCAT in context of leashed and unleashed construct.
(C) Quantification of PI(3)P generated by leashed and unleashed PI3KC3-C1 from PI and [γ-32 P] ATP. Samples were normalized to the activity of wild-type PI3KC3-C1 complex.
(D) Pho8Δ60 assay to monitor autophagy in yeast was carried out in rich (gray) or nitrogen starvation (black) media. Samples were normalized to the activity of Vps34 in rapamycin-treated cells.
(E) Quantification of PI(3)P generated by fully assembled PI3KC3-C1 and as compared to isolated full-length VPS34. Data are represented as mean of three independent experiments ± SD.
See also Figures S1–S4.
Molecular Cell  , e3DOI: ( /j.molcel )
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6. Figure 4 PI3K Complex Assembly and Activation Model
Schematic of the PI3KC3-C1 complex showing the role of VPS15 in the assembly pathway and in regulating the dynamics and activity of the VPS34 HELCAT.
Molecular Cell  , e3DOI: ( /j.molcel )
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