Defects in Galactose Metabolism and Glycoconjugate Biosynthesis in a UDP-Glucose Pyrophosphorylase-Deficient Cell Line Are Reversed by Adding Galactose to the Growth Medium

Autor: Giusy Sala, Riccardo Ghidoni, Monica Thelestam, Stuart E.H. Moore, Christelle Durrant, Jana I. Fuehring, Agnès Rötig, Alexandra Willemetz, Abram Katz, D Chretien, Myriam Ermonval
Přispěvatelé: Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Hannover Medical School [Hannover] (MHH), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Università degli Studi di Milano = University of Milan (UNIMI), Karolinska Institutet [Stockholm], Institut Pasteur [Paris] (IP), Département de Virologie - Department of Virology, The authors’ laboratories were supported by: The Mizutani Foundation, the GIS- Institut des maladies rares/INSERM-funded French CDG Research Network, European Union FP6-Coordination Action EUROGLYCANET (LSHM-CT-2005-512131), the E-Rare-2 Joint Transnational Call 2011 (EURO-CDG), institutional funding from INSERM, La Fondation pour la Recherche Médicale (FRM), Swedish Research Council (grant number 05969)., We thank Eduardo Osinaga (Depto. de Bioquímica, Facultad de Medicina, Montevideo, Uruguay) for supplying anti-Tn MAb 83D4., European Project: 30050,EUROGLYCANET, Ermonval, Myriam, Congenital Disorders of Glycosylation: a European network for the advancement of research, diagnosis and treatment of a growing group of rare disorders - EUROGLYCANET - 30050 - OLD, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Università degli Studi di Milano [Milano] (UNIMI), Institut Pasteur [Paris]
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Glycosylation
Glycoconjugate
MESH: Brain Diseases
MESH: Cricetinae
MESH: Congenital Disorders of Glycosylation
lcsh:Chemistry
chemistry.chemical_compound
Congenital Disorders of Glycosylation
developmental epileptic encephalopathies (dees)
protein glycosylation
Cricetinae
ugp2
MESH: Animals
Lung
lcsh:QH301-705.5
Spectroscopy
chemistry.chemical_classification
Brain Diseases
0303 health sciences
Growth medium
congenital disorders of glycosylation (cdgs)
MESH: Kinetics
glycosphingolipid
UTP—glucose-1-phosphate uridylyltransferase
030302 biochemistry & molecular biology
MESH: Glycosphingolipids
MESH: Uridine Diphosphate Glucose
General Medicine
MESH: Glycosylation
3. Good health
Computer Science Applications
Biochemistry
Uridine Diphosphate Glucose
MESH: Galactose
UTP-Glucose-1-Phosphate Uridylyltransferase
galactose
udp-glucose
MESH: UTP-Glucose-1-Phosphate Uridylyltransferase
[SDV.BC]Life Sciences [q-bio]/Cellular Biology
Article
Glycosphingolipids
Catalysis
Cell Line
Inorganic Chemistry
03 medical and health sciences
Biosynthesis
Extracellular
Animals
Humans
MESH: Lung
Physical and Theoretical Chemistry
Molecular Biology
MESH: Glycoconjugates
[SDV.BC] Life Sciences [q-bio]/Cellular Biology
030304 developmental biology
MESH: Humans
Organic Chemistry
udp-galactose
Culture Media
MESH: Cell Line
carbohydrates (lipids)
Kinetics
chemistry
lcsh:Biology (General)
lcsh:QD1-999
Cell culture
prion protein
Galactose
MESH: Culture Media
Glycoconjugates
Zdroj: International Journal of Molecular Sciences, Vol 21, Iss 6, p 2028 (2020)
International Journal of Molecular Sciences
International Journal of Molecular Sciences, 2020, 21 (6), pp.2028. ⟨10.3390/ijms21062028⟩
Volume 21
Issue 6
International Journal of Molecular Sciences, MDPI, 2020, 21 (6), pp.2028. ⟨10.3390/ijms21062028⟩
ISSN: 1422-0067
1661-6596
Popis: UDP-glucose (UDP-Glc) is synthesized by UGP2-encoded UDP-Glc pyrophosphorylase (UGP) and is required for glycoconjugate biosynthesis and galactose metabolism because it is a uridyl donor for galactose-1-P (Gal1P) uridyltransferase. Chinese hamster lung fibroblasts harboring a hypomrphic UGP(G116D) variant display reduced UDP-Glc levels and cannot grow if galactose is the sole carbon source. Here, these cells were cultivated with glucose in either the absence or presence of galactose in order to investigate glycoconjugate biosynthesis and galactose metabolism. The UGP-deficient cells display <
5% control levels of UDP-Glc/UDP-Gal and >
100-fold reduction of [6-3H]galactose incorporation into UDP-[6-3H]galactose, as well as multiple deficits in glycoconjugate biosynthesis. Cultivation of these cells in the presence of galactose leads to partial restoration of UDP-Glc levels, galactose metabolism and glycoconjugate biosynthesis. The Vmax for recombinant human UGP(G116D) with Glc1P is 2000-fold less than that of the wild-type protein, and UGP(G116D) displayed a mildly elevated Km for Glc1P, but no activity of the mutant enzyme towards Gal1P was detectable. To conclude, although the mechanism behind UDP-Glc/Gal production in the UGP-deficient cells remains to be determined, the capacity of this cell line to change its glycosylation status as a function of extracellular galactose makes it a useful, reversible model with which to study different aspects of galactose metabolism and glycoconjugate biosynthesis.
Databáze: OpenAIRE
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