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Recombinant Human Lysyl Oxidase-like 2 Secreted from Human Embryonic Kidney Cells Displays Complex and Acidic Glycans at All Three N-Linked Glycosylation Sites
journal contribution
posted on 2018-04-05, 00:00 authored by Eden P. Go, Hee-Jung Moon, Minae Mure, Heather DesaireHuman
lysyl oxidase-like 2 (hLOXL2), a glycoprotein implicated
in tumor progression and organ fibrosis, is a molecular target for
anticancer and antifibrosis treatment. This glycoprotein contains
three predicted N-linked glycosylation sites; one
is near the protein’s active site, and at least one more is
known to facilitate the protein’s secretion. Because the glycosylation
impacts the protein’s biology, we sought to characterize the
native, mammalian glycosylation profile and to determine how closely
this profile is recapitulated when the protein is expressed in insect
cells. All three glycosylation sites on the protein, expressed in
human embryonic kidney (HEK) cells, were characterized individually
using a mass spectrometry-based glycopeptide analysis workflow. These
data were compared to the glycosylation profile of the same protein
expressed in insect cells. We found that the producer cell type imparts
a substantial influence on the glycosylation of this important protein.
The more-relevant version, expressed in HEK cells, contains large,
acidic glycoforms; these glycans are not generated in insect cells.
The glycosylation differences likely have structural and functional
consequences, and these data should be considered when generating
protein for functional studies or for high-throughput screening campaigns.
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producer cell typehigh-throughput screening campaignsproteinglycosylation sitesglycosylation profileinsect cellsmass spectrometry-based glycopeptide analysis workflowGlycosylation Sites Human lysyl oxidase-like 2HEKRecombinant Human Lysyl Oxidase-like 2 SecretedHuman Embryonic Kidney Cells Displays Complex
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