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Arabidopsis Wdr8 interactors identification

ABSTRACT: To identify Arabidopsis Wdr8 interactors, immunoprecipitation associated LC-MS/MS analysis was carried out. Crude proteins extracted from Wdr8-GFP expressing transgenic plants was subjected to the immuno-precipitation assay using GFP antibody magnetic beads (µMACS GFP isolation kit, Miltenyi Biotec). Co-purified proteins were separated by 10% SDS-PAGE gel and stained with SYPRO Ruby (BioRad laboratories). The stained protein bands were excised into several fraction pieces according to protein sizes, and in-gel protein digestion by trypsin was carried out. Purified protein peptides were subjected to LC-MS/MS analysis (LTQ-Orbitrap XL-HTC-PAL system). Collected MS/MS peak spectra was analyzed by the MASCOT server to identify peptide sequence.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Seedling

SUBMITTER: Noriyoshi Yagi

LAB HEAD: Takashi Hashimoto

PROVIDER: PXD026063 | Pride | 2022-02-17

REPOSITORIES: Pride

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An anchoring complex recruits katanin for microtubule severing at the plant cortical nucleation sites.

Yagi Noriyoshi N Kato Takehide T Matsunaga Sachihiro S Ehrhardt David W DW Nakamura Masayoshi M Hashimoto Takashi T

Nature communications 20210617 1

Microtubules are severed by katanin at distinct cellular locations to facilitate reorientation or amplification of dynamic microtubule arrays, but katanin targeting mechanisms are poorly understood. Here we show that a centrosomal microtubule-anchoring complex is used to recruit katanin in acentrosomal plant cells. The conserved protein complex of Msd1 (also known as SSX2IP) and Wdr8 is localized at microtubule nucleation sites along the microtubule lattice in interphase Arabidopsis cells. Katan ...[more]

PMID: 34140499

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