Investigating N-terminal shedding of GPR125 in GPR125 overexpressing HEK293T cells using N- and C-terminal enrichment and different cleavage strategies

ABSTRACT: The study was conducted to elucidate potential cleavage sites of GPR125 which are known to undergo N-terminal shedding through autoproteolysis in a region different to other GRPs. To do these different enzymatic strategies as well as enrichment of both N-terminal and C-terminal peptides, was conducted on media from GPR125 overexpressing HEK cells. Other sample types such as media from non-transfected HEK cells and non-PNGnase treated media showed barely any signal from GPR125. GluC was also used as the common clevage site is located in a region devoid of R/K residues. However, few GPR125 originating peptides were identified from these samples compared to Trypsin and TrypN treated samples. Lysate was also briefly analysed resulting in peptides spanning more of the C-terminal end of the protein which were not previously found in the up-concentrated media.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: Nicoline Wichmand Thorsen  

LAB HEAD: Michael J Davies

PROVIDER: PXD083139 | Pride | 2026-08-31

REPOSITORIES: Pride