Comparison of two expression systems using COS7 cells and yeast cells for expression of heart/muscle-type carnitine palmitoyltransferase 1

Unique Behavior of Bacterially Expressed Rat Carnitine Palmitoyltransferase 2 and Its Catalytic Activity

“…Finally, we asked whether bacterially expressed full-length CPT2, partially purified by simple centrifugation as described above, shows catalytic activity. As a comparison, we also expressed rat CPT1b in COS7 cells as previously described, [14][15][16] and used lysates of these cells to represent CPT1b. As shown in Fig.…”

“…L-[methyl-3 H] carnitine hydrochloride as a tracer, essentially as described previously for measurement of CPT1 activity. [14][15][16] Briefly, the reaction was initiated by addition of 50 µL sample solution containing 30 µg protein sample of crudely purified CPT2 to the prewarmed reaction solution (450 µL), to make final concentrations of 150 mM KCl, 1 mM EDTA, 0.25 mM reduced glutathione, 1.3 mg/mL fatty acid-free bovine serum albumin (BSA), 0.5 mM L-carnitine, 50 µM palmitoyl-CoA, 2 mM NaCN, 9.25 kBq L-[methyl-3 H] carnitine and 50 mM N-(2-hydroxyethyl) piperazine-N′-2-ethanesulfonic acid (Hepes) buffer, pH 7.4. After incubation at 30 °C for 20 min, the reaction was terminated by the addition of 200 µL 3 M HC1.…”

Dioxygen and Metabolism; Dangerous Liaisons in Cardiac Function and Disease

“…For long-chain FA, a carnitine shuttle is required, facilitated by the carnitine-palmitoyl transferase 1 (CPT1) (Saggerson, 1982 ; Bonnefont et al, 2004 ). CPT1B is the major isoform located at the outer membrane of the cardiomyocyte mitochondria (Saggerson, 1982 ; Leij et al, 2002 ; Hada et al, 2012 ). In the heart, CPT1B represents a checkpoint that responds to metabolic feedback for FAO (He et al, 2012 ).…”

PHDs/CPT1B/VDAC1 axis regulates long-chain fatty acid oxidation in cardiomyocytes

“…Located at the outer mitochondrial membrane (OMM), carnitine O-palmitoyltransferase 1b (CPT1B), the major CPT1 isoform expressed in heart, catalyzes the carnitine conjugation of LCFAs to facilitate their mitochondria uptake and plays a key role in LCFA β-oxidation in cardiomyocytes ( Declercq et al, 1987 ; Hada et al, 2012 ; van der Leij et al, 2002 ). Although no disease-causing mutation has been reported in humans, CPT1B depletion in striated muscles leads to a fatal hypertrophic cardiomyopathy associated with lipotoxicity in mouse ( Haynie et al, 2014 ).…”