Proteomics

Dataset Information

0

N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris


ABSTRACT: 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.

INSTRUMENT(S): LTQ Orbitrap, Q Exactive

ORGANISM(S): Homo Sapiens (human) Komagataella Pastoris (yeast) (pichia Pastoris)

TISSUE(S): Cell Suspension Culture, Vegetative Cell (sensu Fungi)

DISEASE(S): Disease Free

SUBMITTER: Sergo Kasvandik  

LAB HEAD: Nigulas Samel

PROVIDER: PXD000965 | Pride | 2014-08-18

REPOSITORIES: Pride

altmetric image

Publications

N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris.

Kukk Kaia K   Kasvandik Sergo S   Samel Nigulas N  

SpringerPlus 20140815


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. Correct N-glycosylation is proposed to be obligatory for proper folding of mammalian PGHSs. In this study, human PGHS-1 and -2 (hPGHS-1 and -2) were expressed in the yeast Pichia pastoris. Reco  ...[more]

Similar Datasets

2021-03-15 | PXD024672 | Pride
2015-11-20 | MODEL1510220000 | BioModels
2023-04-23 | PXD039002 | Pride
2022-11-02 | PXD037229 | Pride
2011-04-29 | GSE28920 | GEO
2017-09-20 | PXD007267 | Pride
2014-12-08 | PXD001284 | Pride
2017-08-08 | PXD005708 | Pride
2023-08-09 | GSE239877 | GEO
| E-GEOD-28920 | biostudies-arrayexpress