Studies of porphyrin synthesis in fibroblasts of patients with congenital erythropoietic porphyria and one patient with homozygous coproporphyria

Successful therapeutic effect in a mouse model of erythropoietic protoporphyria by partial genetic correction and fluorescence-based selection of hematopoietic cells

“…Ferrochelatase activity was measured in the BM and liver according to Camadro and Labbe. 34 Protoporphyrin levels in RBC, plasma and organs were determined spectrofluorimetrically 35 using a Hitachi F-4500 fluorescence spectrophotometer (Braun Sciencetec, Les Ulis, France) (excitation 405 nm and emission 605 nm), after extraction with 1 mol/l HCLO 4 /CH 3 OH (1:1, v/v). Percentage of fluorescent RBC was scored at the end of the study.…”

The mitochondrial location of protoporphyrinogen oxidase

“…One was diluted 30 times in a medium containing 100 mM Tris/Cl buffer pH 8.7, 2 mM dithiothreitol, 1 mM EDTA and 0.1% Triton X-100 (v/v), and protoporphyrin formation estimated fluorimetrically as described for assay of protoporphyrinogen oxidase activity; the second aliquot was diluted 40 times in ethyl acetate/acetic acid mixture (4: 1, v/v) and exposed to light for 2 h at room temperature in order to ensure complete oxidation of porphyrinogens to corresponding porphyrins. Protoporphyrinogen and protoporphyrin formed were then determined fluorimetrically as total oxidized protoporphyrin [13]. Protoporphyrinogen was determined from the difference between total protoporphyrin measured after reoxidation in ethyl acetate/acetic acid mixture and protoporphyrin formation determined directly after incubation with coproporphyrinogen.…”

Reversion of hepatobiliary alterations by bone marrow transplantation in a murine model of erythropoietic protoporphyria

“…Determination of ferrochelatase activity was performed according to Camadro, et al 21 Ferrochelatase activity was expressed as nmol of meso-Zn formed per hour per mg of protein (U/mg). Protoporphyrin concentration was determined spectrofluorometrically 22 using a Hitachi F-4500 fluorescence spectrophotometer (Braun Sciencetec, Les Ulis, France).…”