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BioID2_PHB2(V1)-(WT_KKK_I225P)

ABSTRACT: HEK293 cells that stably express wild-type (WT), E229/231/233K (KKK), or I225P mutant PHB2(V1) fused at C-terminus with the biotin ligase from A. aeolicus were incubated with 100 μM biotin for 18 h. The mitochondrial fractions were lysed in RIPA buffer, and the clarified supernatants were incubated with streptavidin-agarose beads for 2 h at 4° C. Proteins on the beads were digested by adding trypsin/Lys-C mix for 16 h at 37° C and analyzed by LC-MS/MS. Label-free quantification was performed using Proteome Discoverer 2.2 software.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Hidetaka Kosako

PROVIDER: PXD011946 | JPOST Repository | Thu Sep 05 00:00:00 BST 2019

REPOSITORIES: jPOST

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Structural Basis of Mitochondrial Scaffolds by Prohibitin Complexes: Insight into a Role of the Coiled-Coil Region.

Yoshinaka Takahiro T Kosako Hidetaka H Yoshizumi Takuma T Furukawa Ryo R Hirano Yu Y Kuge Osamu O Tamada Taro T Koshiba Takumi T

iScience 20190903

The coiled-coil motif mediates subunit oligomerization and scaffolding and underlies several fundamental biologic processes. Prohibitins (PHBs), mitochondrial inner membrane proteins involved in mitochondrial homeostasis and signal transduction, are predicted to have a coiled-coil motif, but their structural features are poorly understood. Here we solved the crystal structure of the heptad repeat (HR) region of PHB2 at 1.7-Å resolution, showing that it assembles into a dimeric, antiparallel coil ...[more]

PMID: 31522117

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