Universität zu Köln

Janes, Verena, Grabany, Simona, Delbrouck, Julien, Vincent, Stephane P., Gottschalk, Johannes ORCID: 0000-0002-7556-3663, Elling, Lothar ORCID: 0000-0002-3654-0397 and Hanisch, Franz-Georg (2020). Fluorinated Galactoses Inhibit Galactose-1-Phosphate Uridyltransferase and Metabolically Induce Galactosemia-like Phenotypes in HEK-293 Cells. Cells, 9 (3). BASEL: MDPI. ISSN 2073-4409

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Abstract

Genetic defects of human galactose-1-phosphate uridyltransferase (hGALT) and the partial loss of enzyme function result in an altered galactose metabolism with serious long-term developmental impairment of organs in classic galactosemia patients. In search for cellular pathomechanisms induced by the stressor galactose, we looked for ways to induce metabolically a galactosemia-like phenotype by hGALT inhibition in HEK293 cells. In kinetic studies, we provide evidence for 2-fluorinated galactose-1-phosphate (F-Gal-1-P) to competitively inhibit recombinant hGALT with a K-I of 0.9 mM. Contrasting with hepatic cells, no alterations of N-glycoprofiles in MIG (metabolic induction of galactosemia)-HEK293 cells were revealed for an inducible secretory netrin-1 probe by MALDI-MS. Differential fluorescence-activated cell sorting demonstrated reduced surface expression of N-glycosylated CD109, EGFR, DPP4, and rhMUC1. Membrane raft proteomes exhibited dramatic alterations pointing to an affection of the unfolded protein response, and of targeted protein traffick. Most prominent, a negative regulation of oxidative stress was revealed presumably as a response to a NADPH pool depletion during reduction of Gal/F-Gal. Cellular perturbations induced by fluorinated galactoses in normal epithelial cells resemble proteomic changes revealed for galactosemic fibroblasts. In conclusion, the metabolic induction of galactosemia-like phenotypes in healthy epithelial/neuronal cells could support studies on the molecular pathomechanisms in classic galactosemia, in particular under conditions of low galactose stress and residual GALT activity.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Janes, Verena UNSPECIFIED UNSPECIFIED UNSPECIFIED Grabany, Simona UNSPECIFIED UNSPECIFIED UNSPECIFIED Delbrouck, Julien UNSPECIFIED UNSPECIFIED UNSPECIFIED Vincent, Stephane P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Gottschalk, Johannes UNSPECIFIED orcid.org/0000-0002-7556-3663 UNSPECIFIED Elling, Lothar UNSPECIFIED orcid.org/0000-0002-3654-0397 UNSPECIFIED Hanisch, Franz-Georg UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-342138
DOI:	10.3390/cells9030607
Journal or Publication Title:	Cells
Volume:	9
Number:	3
Date:	2020
Publisher:	MDPI
Place of Publication:	BASEL
ISSN:	2073-4409
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language URINARY EXOVESICLES; N-GLYCOSYLATION; O-GLYCAN; FLUOROPHOSPHORYLATION; PROTEOMICS; TOR Multiple languages Cell Biology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/34213