PXD024117-1

PXD024117 is an original dataset announced via ProteomeXchange.

Title	Endogenous THBS1 is C-mannosylated by human induced pluripotent stem cells.
Description	We used a human thrombospondin-1 (THBS1) fragment comprising thrombospondin type 1 repeats (TSRs) 1 to 3, which was recombinantly expressed in HEK 293T cells and digested with AspN to determine chromatographic retention times and to optimise MS parameters. We detected two peptides from TSR2 and TSR3 containing the consensus site for C-mannosylation in their non-, mono- and di-C-mannosylated state. These peptides – irrespective of C-mannosylation - were detected at less than 3 % of the intensity of two other THBS-derived peptides which do not contain a WxxW motif (Figure 3A). With the knowledge of the specific chromatographic retention times and the intensity patterns of fragment ions for the two peptides containing the WxxW sites for C-mannosylation from the recombinant THBS1, we now analysed a native sample of hiPSC-derived proteins from cell culture supernatants by MRM. Indeed we detected the most intense fragment ions for the peptide DACPINGGWGPWSPW with one or two C-mannoses at the expected elution times and with the expected intensity profiles (Figure 3B).
HostingRepository	PanoramaPublic
AnnounceDate	2021-05-12
AnnouncementXML	Submission_2021-05-12_11:02:38.614.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Roman Sakson
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Hex; Carbamidomethyl
Instrument	QTRAP 5500

Revision	Datetime	Status	ChangeLog Entry
0	2021-02-10 09:04:15	ID requested
⏵ 1	2021-05-12 11:02:39	announced

Cirksena K, H, ü, tte HJ, Shcherbakova A, Thumberger T, Sakson R, Weiss S, Jensen LR, Friedrich A, Todt D, Kuss AW, Ruppert T, Wittbrodt J, Bakker H, Buettner FFR, The C-Mannosylome of Human Induced Pluripotent Stem Cells Implies a Role for ADAMTS16 C-Mannosylation in Eye Development. Mol Cell Proteomics, 20():100092(2021) [pubmed]

submitter keyword: hiPSC, TSR, thrombospondin, C-Mannosylation

Falk F. R. Buettner
contact affiliation	Institute of Clinical Biochemistry, Hannover Medical School, 30625 Hannover, Germany
contact email	Buettner.Falk@mh-hannover.de
lab head
Roman Sakson
contact affiliation	Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany
contact email	roman.sakson@gmx.net
dataset submitter

Panorama Public dataset URI