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PXD018626

PXD018626 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleAntibody-free targeted proteomics assay for absolute measurement of α-tubulin acetylation
DescriptionAcetylation 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.
HostingRepositoryPanoramaPublic
AnnounceDate2020-07-30
AnnouncementXMLSubmission_2020-07-30_05:33:05.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterAlok Shah
SpeciesList scientific name: Mus musculus; NCBI TaxID: 10090; scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationListAcetyl; Carbamidomethyl; Label:13C(6)15N(1)
Instrument6490 Triple Quadrupole LC/MS
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-04-17 07:19:34ID requested
12020-07-30 05:33:06announced
Publication List
Shah AK, Wali G, Sue CM, Mackay-Sim A, Hill MM, Antibody-free targeted proteomics assay for absolute measurement of α-tubulin acetylation. Anal Chem, ():(2020) [pubmed]
Keyword List
submitter keyword: α-tubulin acetylation
Contact List
Michelle Hill
contact affiliationThe University of Queensland and QIMR Berghofer Medical Research Institute, Brisbane, Australia
contact emailmichelle.hill@qimrberghofer.edu.au
lab head
Alok Shah
contact affiliationQIMR Berghofer Medical Research Institute, Brisbane, Australia
contact emailalok.shah@qimrberghofer.edu.au
dataset submitter
Full Dataset Link List
Panorama Public dataset URI