PXD030550

PXD030550 is an original dataset announced via ProteomeXchange.

Title	Glycoproteoforms of Integrins α2 and β1 in Megakaryocytes in the Occurrence of Primary Myelofibrosis
Description	Primary myelofibrosis (PMF) is a myeloproliferative neoplasm prone to leukemic transformation, for which limited treatment is available. Amongst individuals diagnosed with PMF, the most prevalent mutation is the JAK2V617F somatic point mutation that activates the Janus kinase 2 (JAK2) enzyme. Our earlier reports on hyperactivity of β1 integrin and enhanced adhesion activity of the α2β1 complex in JAK2V617F megakaryocytes (MKs) led us to examine the new hypothesis that this mutation leads to post-translational modification via changes in glycosylation. Samples were derived from immunoprecipitation of MKs obtained from Vav1-hJAK2V617F and wild type mice. Immunoprecipitated fractions were separated by SDS-PAGE and analyzed using LC-MS/MS techniques in a bottom-up glycoproteomics workflow. In the immunoprecipitate, glycopeptiforms corresponding to 11 out of the 12 potential N-glycosylation sites of integrin β1, and to all nine potential glycosylation sites of integrin α2, were observed. Glycopeptiforms were compared across WT and JAK2V617F phenotypes for both integrins. The overall trend observed is that JAK2V617F mutation in PMF MKs leads to changes in β1 glycosylation; in most cases, it results in an increase in the integrated area of glycopeptiforms. We also observed that, in mutated MKs, changes in integrin α2 glycosylation were more substantial than those observed for integrin β1 glycosylation, a finding that suggests that altered integrin α2 glycosylation may also affect activation. Additionally, the identification of proteins associated to the cytoskeleton that were co-immunoprecipitated with integrins α2 and β1 demonstrated the potential of the methodology employed in this study to provide some insight, at the peptide level, into the consequences of integrin activation in MKs. The extensive and detailed glycosylation patterns we uncovered provide a basis for future functional studies of each site in control cells as compared to JAK2V617F mutated cells.
HostingRepository	PRIDE
AnnounceDate	2022-02-22
AnnouncementXML	Submission_2022-02-22_04:19:15.344.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD030550
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	MAISSA GAYE
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Mesocricetus auratus (Golden hamster); NCBI TaxID: 10036;
ModificationList	complex glycosylation
Instrument	Orbitrap Fusion Lumos; Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-20 11:17:26	ID requested
⏵ 1	2022-02-22 04:19:16	announced

Gaye MM, Ward CM, Piasecki AJ, Stahl VL, Karagianni A, Costello CE, Ravid K, 1 in Megakaryocytes in the Occurrence of JAK2V617F Mutation-Induced Primary Myelofibrosis. Mol Cell Proteomics, 21(4):100213(2022) [pubmed]

submitter keyword: Integrins

megakaryocytes

primary myelofibrosis

LC-MS/MS

glycopeptiforms

Catherine E. Costello
contact affiliation	Boston University, School of Medicine
contact email	cecmsms@bu.edu
lab head
MAISSA GAYE
contact affiliation	Boston University School of Medicine
contact email	gayem@bu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD030550
     1. Label: PRIDE project
     2. Name: Glycoproteoforms of Integrins α2 and β1 in Megakaryocytes in the Occurrence of Primary Myelofibrosis