PXD000965

PXD000965 is an original dataset announced via ProteomeXchange.

Title	N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris
Description	Prostaglandin H synthases (PGHSs) are N-glycosylated membrane proteins that catalyse the committed step in prostaglandin synthesis. Unlike PGHS-2, the production of recombinant PGHS-1 in non-mammalian expression systems is complicated. The majority of the heterologous enzyme is inactive due to misfolding. N-glycosylation is proposed to be obligatory for the correct folding of mammalian PGHSs. In this study, human PGHS-1 and -2 (hPGHS-1 and -2) were expressed in the yeast Pichia pastoris, and the N-glycosylation patterns of the purified recombinant proteins were characterised using nano-LC/MS/MS. Recombinant hPGHS-2 was catalytically active, whereas hPGHS-1 was inactive. Unexpectedly, the accumulation of non-glycosylated hPGHS-1 was not observed in the crude lysate of the yeast cells. In addition, the purified hPGHS isoforms exhibited similar N-glycosylation site occupancy. The results indicate that there are more complex grounds for the inactivity of the recombinant hPGHS-1 produced in yeast.
HostingRepository	PRIDE
AnnounceDate	2014-08-18
AnnouncementXML	Submission_2014-09-10_01:29:34.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sergo Kasvandik
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Komagataella pastoris (Yeast) (Pichia pastoris); NCBI TaxID: 4922;
ModificationList	deamidated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue; (18)O label at both C-terminal oxygens; 1x(18)O labeled deamidated L-asparagine
Instrument	LTQ Orbitrap; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2014-05-09 05:44:46	ID requested
1	2014-08-18 00:37:25	announced
⏵ 2	2014-09-10 01:29:35	announced	Updated publication reference for PubMed record(s): 25170432.

Kukk K, Kasvandik S, Samel N, N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris. Springerplus, 3():436(2014) [pubmed]

curator keyword: Biomedical

submitter keyword: recombinant protein,PGHS, N-glycosylation, proteomics, LC-MS/MS

Nigulas Samel
contact affiliation	Tallinn University of Technology
contact email	nigulas.samel@ttu.ee
lab head
Sergo Kasvandik
contact affiliation	University of Tartu, Proteomics core facility
contact email	sergo.kasvandik@ut.ee
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD000965
     1. Label: PRIDE project
     2. Name: N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris