PXD018626

PXD018626 is an original dataset announced via ProteomeXchange.

Title	Antibody-free targeted proteomics assay for absolute measurement of α-tubulin acetylation
Description	Acetylation of α-tubulin at conserved lysine 40 (K40) amino acid residue regulates microtubule dynamics and controls a wide range of cellular activities. Dysregulated microtubule dynamics characterised by differential α-tubulin acetylation is a hallmark of cancer, neurodegeneration and other complex disorders. Hence, accurate quantitation of α-tubulin acetylation is required in human disease and animal model studies. We developed a novel antibody-free proteomics assay to measure α-tubulin acetylation targeting protease AspN-generated peptides harbouring K40 site. Using the synthetic unmodified and acetylated stable isotope labelled peptides DKTIGGG and DKTIGGGD, we demonstrate assay linearity across 4 log magnitude and reproducibility of <10% coefficient of variation . The assay accuracy was validated by titration of 10% to 80% mixture of acetylated/non-acetylated α-tubulin peptides in the background of human olfactory neurosphere-derived stem (ONS) cell matrix. Furthermore, in agreement with antibody-based high content microscopy analysis, the targeted proteomics assay reported an induction of α-tubulin K40 acetylation upon Trichostatin A stimulation of ONS cells. Independently, we found 35.99% and 16.11% α-tubulin acetylation for mouse spinal cord and brain homogenate tissue, respectively, as measured by our assay. In conclusion, this simple, antibody-free proteomics assay enables quantitation of α-tubulin acetylation, and is applicable across various fields of biology and medicine.
HostingRepository	PanoramaPublic
AnnounceDate	2020-07-30
AnnouncementXML	Submission_2020-07-30_05:33:05.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alok Shah
SpeciesList	scientific name: Mus musculus; NCBI TaxID: 10090; scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Acetyl; Carbamidomethyl; Label:13C(6)15N(1)
Instrument	6490 Triple Quadrupole LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-17 07:19:34	ID requested
⏵ 1	2020-07-30 05:33:06	announced

Shah AK, Wali G, Sue CM, Mackay-Sim A, Hill MM, -Tubulin Acetylation. Anal Chem, 92(16):11204-11212(2020) [pubmed]

submitter keyword: α-tubulin acetylation

Michelle Hill
contact affiliation	The University of Queensland and QIMR Berghofer Medical Research Institute, Brisbane, Australia
contact email	michelle.hill@qimrberghofer.edu.au
lab head
Alok Shah
contact affiliation	QIMR Berghofer Medical Research Institute, Brisbane, Australia
contact email	alok.shah@qimrberghofer.edu.au
dataset submitter

Panorama Public dataset URI