Hyperinsulinism associated with GLUD1 mutation: allosteric regulation and functional characterization of p.G446V glutamate dehydrogenase

摘要

Mitochondrial metabolism in EBV-transformed lymphoblasts. The metabolic function of LCL lymphoblasts derived from control subject (GDH-wt) or from patient with mutant GDH (GDH-G446V) assessed using a 96-well plate Seahorse analyzer. Extracellular acidification rate (ECAR, ) and the oxygen consumption rate (OCR, ) were measured under the stimulation with 5 mM alanine and 5 mM glutamine, followed by the sequential addition of oligomycin (O), FCCP (F), and antimycin A/rotenone (A/R). ATP production ( ) and maximal respiration ( ) were calculated from OCR raw data. Values are shown as means ± SD, = 10, * < 0.05, ** < 0.01, *** < 0.001 for wild-type GDH-wt versus GDH-G446V