Dataset Information

Proteins detected in HA-Mmr1 IP fraction; In vitro Mmr1 phosphorylation

ABSTRACT: A capillary reverse-phase HPLC-MS/MS system was composed of a Dionex U3000 gradient pump equipped with a VICI Cheminert valve and Q Exactive equipped with a nano-electrospray ionization (NSI) source (AMR, Tokyo, Japan). The desalted peptides were loaded into a separation capillary C18 reverse-phase column (NTCC-360/100-3-125, 125×0.1 mm, Nikkyo Technos, Tokyo, Japan). Xcalibur 3.0.63 system (Thermo) was used to record peptide spectra over the mass range of m/z 350–1800. Repeatedly, MS spectra were recorded, followed by ten data-dependent high energy collisional dissociation (HCD) MS/MS spectra generated from the ten highest intensity precursor ions. MS/MS spectra were interpreted, and peak lists were generated using Proteome Discoverer 2.2.0.388 (Thermo). Searches were performed using the SEQUEST (Thermo) program against Saccharomyces cerevisiae (SwissProt TaxID = 559292) peptide sequence.

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Takumi Kamura

PROVIDER: PXD032866 | JPOST Repository | Tue Mar 29 00:00:00 BST 2022

REPOSITORIES: jPOST

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20180611_QE2_HTSPAL_sand_AcOH_PE_2018-150_e6.raw	Raw
	20200403_QE1_HTCPAL_sand_AcOH_PE_SUSValve_2020-079_e1-A.raw	Raw
	20200403_QE1_HTCPAL_sand_AcOH_PE_SUSValve_2020-079_e2.raw	Raw
	20200403_QE1_HTCPAL_sand_AcOH_PE_SUSValve_2020-079_e3-A.raw	Raw
	InVitroP2020-079_20200403_QE1_HTCPAL_sand_AcOH_PE_SUSValve_2020-079_all_MMR1_2Slim.xlsx	Xlsx

Items per page:

1 - 5 of 6

Publications

Proteolysis of adaptor protein Mmr1 during budding is necessary for mitochondrial homeostasis in Saccharomyces cerevisiae.

Obara Keisuke K Yoshikawa Taku T Yamaguchi Ryu R Kuwata Keiko K Nakatsukasa Kunio K Nishimura Kohei K Kamura Takumi T

Nature communications 20220414 1

In yeast, mitochondria are passed on to daughter cells via the actin cable, motor protein Myo2, and adaptor protein Mmr1. They are released from the actin-myosin machinery after reaching the daughter cells. We report that Mmr1 is rapidly degraded by the ubiquitin-proteasome system in Saccharomyces cerevisiae. Redundant ubiquitin ligases Dma1 and Dma2 are responsible for Mmr1 ubiquitination. Dma1/2-mediated Mmr1 ubiquitination requires phosphorylation, most likely at S414 residue by Ste20 and Cla ...[more]

PMID: 35422486

Similar Datasets

Project description:Ten human ovarian cancer cell lines were kindly provided as follows: an ovarian serous adenocarcinoma cell line, SKOV3 (Dr. N. Nagai, Hiroshima University, Hiroshima, Japan); KF28 and its TXL/CDDP-resistant variant (Dr. Y. Kikuchi, National Defense Medical College, Saitama, Japan); KF, SHIN3 and 5 ovarian clear cell adenocarcinoma cell lines, KK, TAYA, RMG-1, OVISE and OVTOKO (Dr. M. Suzuki, Jichi Medical School, Tochigi, Japan). All cell lines were cultured in RPMI 1640 medium (Life Technologies, Inc., Grand Island, NY) containing 10% heat-inactivated fetal bovine serum (FBS; BioWhittaker, Verviers, Belgium) in a humidified atmosphere of 5% CO2 and maintained in continuous exponential growth by passage every 3 days. Exponentially growing cultured cells were collected after two-washings with PBS. The cell pellets were immediately frozen in liquid nitrogen, and stored until use. Total RNA was extracted using QIAGEN RNeasy mini kit (QIAGEN, Inc., Valencia, CA). The quality of the RNA was checked using Agilent Technologies 2100 Bioanalyzer (Agilent, Palo Alto, CA). CodeLink Expression Bioarray System (Amersham Bioscience, Tokyo, Japan) was used according to the manufacturer’s protocol. Briefly, first-strand cDNA was generated from 1 microgram of total RNA of each cell line using reverse transcriptase and a T7 primer, and then second-strand cDNA was produced using DNA polymerase mix and RNase H. Complementary RNA (cRNA) was generated via an in vitro transcription reaction using T7 RNA polymerase and biotin-11-UTP (Perkin Elmer, Boston, MA), which was quantified by spectrometry and checked using Agilent Technologies 2100 Bioanalyzer (Agilent). Ten-microgram of cRNA was then fragmented and hybridized to a Uniset Human 20K I Bioarray containing 19,881 probes with positive and negative bacterial control probes. After hybridization, the arrays were rinsed and labeled with Streptavidin-Cy5, scanned using Agilent DNA Microarray Scanner (Agilent), then analyzed with CodeLink Expression Analysis Software ver.2.3. Expression levels were normalized to the median expression value of the whole array spots. Keywords: parallel sample