PXD036367

PXD036367 is an original dataset announced via ProteomeXchange.

Title	Understanding the role of yeast Yme1 in mitochondrial finction using biochemical and proteomics analysis
Description	Mitochondrial proteome comparison between S. cerevisiae BY4741 WT and Yme1 deletion mutant. Mitochondria were isolated by fractionation as described by Titorenko et al. (2009). Briefly, cells were grown in YPGal and harvested at later exponential phases of OD ~1.5. After pre- treatment with DTT buffer (100 mM Tris-H 2 SO 4 pH 9.4, 10 mM DTT) at 30°C for 20 minutes, the cell wall was digested with zymolyase 20T (Seikagaku Corporation, Japan) in zymolyase buffer (20mM potassium phosphate buffer pH 7.4, 1.2M sorbitol) for 1 hour at 30˚ with shaking at 70 rpm. The resulting spheroblasts were checked under a light microscope to confirm cell wall digestion. Then, the spheroblasts were lysed in homogenization buffer (20mM HEPES-KOH pH7.4, 0.6M sorbitol, 1mM phenylmethylsulfonyl fluoride (PMSF)) on- ice using a glass Teflon homogenizer. The resulting spheroblasts were subjected to two centrifugation steps at 1500 ×g to remove the cell debris and nuclei. Crude mitochondrial materials were isolated from the supernatant by centrifugation at 12,000 ×g at 4˚C for 10 minutes, then resuspended in SEM buffer (250mM sucrose, 0.6M Sorbitol, 20mM HEPES- KOH pH7.4). Protein concentration were checked by measuring absorbance at 280 nm, assuming an absorbance value of A280=0.21 would equate to 10 mg/ml of crude mitochondrial extract. After adjusting to final concentration of 10 mg/ml the crude mitochondria were then flash frozen and stored at −80°C.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:58:18.112.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kwan Ting Kan
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-28 03:25:39	ID requested
1	2022-11-11 03:30:00	announced
⏵ 2	2023-11-14 08:58:18	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: Mitochondria,Yeast, LS/MSMS

Dr. Hui Lu
contact affiliation	School of Biological Sciences; Faculty of Biology, Medicine and Health; The University of Manchester
contact email	hui.lu@manchester.ac.uk
lab head
Kwan Ting Kan
contact affiliation	University of Manchester
contact email	kwanting.kan@manchester.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD036367
     1. Label: PRIDE project
     2. Name: Understanding the role of yeast Yme1 in mitochondrial finction using biochemical and proteomics analysis