Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1977 Aug;60(2):380–389. doi: 10.1172/JCI108787

Comparative Study of Protoporphyrins in Erythropoietic Protoporphyria and Griseofulvin-Induced Murine Protoporphyria

BINDING AFFINITIES, DISTRIBUTION, AND FLUORESCENCE SPECTRA IN VARIOUS BLOOD FRACTIONS

Maureen B Poh-Fitzpatrick 1,2, Angelo A Lamola 1,2, Gregory L Zalar 1,2, Mark Weinstein 1,2, Frank Doleiden 1,2, Maria Freeman 1,2
PMCID: PMC372379  PMID: 874098

Abstract

Excess erythrocyte protoporphyrins of human congenital erythropoietic protoporphyria and of griseofulvin-induced murine hepatic protoporphyria were found to be associated with hemoglobin and stroma fractions in similar relationships. More than 99.5% of total erythrocyte protoporphyrin was bound to hemoglobin in each case. However, profound differences were found when protoporphyrin concentration was measured in erythrocytes that had been segregated into populations of progressive age on discontinuous density gradients. In erythropoietic protoporphyria, porphyrin content diminished rapidly with age; in murine protoporphyria, the aging erythrocyte populations became progressively more porphyrin rich. In vitro diffusion of protoporphyrin from plasma across the intact erythrocyte membrane was demonstrated. The equimolar binding affinity of protoporphyrin to hemoglobin was shown to be 40 times that of protoporphyrin to serum albumin. This strong affinity provides the driving force for the observed transmembrane diffusion, and explains the high erythrocyte/plasma porphyrin ratio in murine hepatic protoporphyria. The opposite rapid efflux of intra-erythrocytic protoporphyrin into plasma previously shown in uncomplicated erythropoietic protoporphyria occurs despite this strong hemoglobin affinity, implying continuous efficient clearance of protoporphyrin from plasma by the liver. Furthermore, these and other data suggest that a hepatic synthetic source for any significant fraction of the blood protoporphyrin in erythropoietic protoporphyria is highly improbable.

Full text

PDF
380

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Corash L. M., Piomelli S., Chen H. C., Seaman C., Gross E. Separation of erythrocytes according to age on a simplified density gradient. J Lab Clin Med. 1974 Jul;84(1):147–151. [PubMed] [Google Scholar]
  2. Cripps D. J., MacEachern W. N. Hepatic and erythropoietic protoporphyria. Delta-aminolevulinic acid synthetase, fluorescence, and microfluorospectrophotometric study. Arch Pathol. 1971 Jun;91(6):497–505. [PubMed] [Google Scholar]
  3. Cripps D. J., Peters H. A. Fluorescing erythrocytes and porphyrin screening tests on urine, stool, and blood. Investigation of photosensitivity. Arch Dermatol. 1967 Dec;96(6):712–720. [PubMed] [Google Scholar]
  4. Cripps D. J., Scheuer P. J. Hepatobiliary changes in erythropoietic protoporphyria. Arch Pathol. 1965 Nov;80(5):500–508. [PubMed] [Google Scholar]
  5. DE MATTEIS F., RIMINGTON C. Disturbance of porphyrin metabolism caused by griseofulvin in mice. Br J Dermatol. 1963 Mar;75:91–104. doi: 10.1111/j.1365-2133.1963.tb13945.x. [DOI] [PubMed] [Google Scholar]
  6. FLODIN P., KILLANDER J. Fractionation of human-serum proteins by gel filtration. Biochim Biophys Acta. 1962 Oct 8;63:402–410. doi: 10.1016/0006-3002(62)90104-x. [DOI] [PubMed] [Google Scholar]
  7. Gschnait F., Konrad K., Hönigsmann H., Denk H., Wolff K. Mouse model for protoporphyria. I. The liver and hepatic protoporphyrin crystals. J Invest Dermatol. 1975 Sep;65(3):290–299. doi: 10.1111/1523-1747.ep12598357. [DOI] [PubMed] [Google Scholar]
  8. Janousek V. The early changes in experimental porphyria of mice induced by griseofulvin. Trans St Johns Hosp Dermatol Soc. 1970;56(1):14–18. [PubMed] [Google Scholar]
  9. Koskelo P., Toivonen I., Rintola P. The binding of 14C-labelled porphyrins by plasma proteins. Clin Chim Acta. 1970 Sep;29(3):559–565. doi: 10.1016/0009-8981(70)90028-8. [DOI] [PubMed] [Google Scholar]
  10. LEWIS G. P. METHOD USING ORTHO-TOLIDINE FOR THE QUANTITATIVE DETERMINATION OF HAEMOGLOBIN IN SERUM AND URINE. J Clin Pathol. 1965 Mar;18:235–239. doi: 10.1136/jcp.18.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lamola A. A., Piomelli S., Poh-Fitzpatrick M. G., Yamane T., Harber L. C. Erythropoietic protoporphyria and lead intoxication: the molecular basis for difference in cutaneous photosensitivity. II. Different binding of erythrocyte protoporphyrin to hemoglobin. J Clin Invest. 1975 Dec;56(6):1528–1535. doi: 10.1172/JCI108234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MAGNUS I. A., JARRETT A., PRANKERD T. A., RIMINGTON C. Erythropoietic protoporphyria. A new porphyria syndrome with solar urticaria due to protoporphyrinaemia. Lancet. 1961 Aug 26;2(7200):448–451. doi: 10.1016/s0140-6736(61)92427-8. [DOI] [PubMed] [Google Scholar]
  13. Marecek Z., Jirsa M., Korínek J. The binding of protoporphyrin to serum proteins. Clin Chim Acta. 1973 May 30;45(4):409–413. doi: 10.1016/0009-8981(73)90042-9. [DOI] [PubMed] [Google Scholar]
  14. Matilla A., Molland E. A. A light and electron microscopic study of the liver in case of erythrohepatic protoporphyria and in griseofulvin-induced porphyria in mice. J Clin Pathol. 1974 Sep;27(9):698–709. doi: 10.1136/jcp.27.9.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakao K., Wada O., Takaku F., Sassa S., Yano Y., Urata G. The origin of the increased protoporphyrin in erythrocytes of mice with experimentally induced porphyria. J Lab Clin Med. 1967 Dec;70(6):923–932. [PubMed] [Google Scholar]
  16. Piomelli S., Davidow B., Guinee V. F., Young P., Gay G. The FEP (free erythrocyte porphyrins) test: a screening micromethod for lead poisoning. Pediatrics. 1973 Feb;51(2):254–259. [PubMed] [Google Scholar]
  17. Piomelli S., Lamola A. A., Poh-Fitzpatrick M. F., Seaman C., Harber L. C. Erythropoietic protoporphyria and lead intoxication: the molecular basis for difference in cutaneous photosensitivity. I. Different rates of disappearance of protoporphyrin from the erythrocytes, both in vivo and in vitro. J Clin Invest. 1975 Dec;56(6):1519–1527. doi: 10.1172/JCI108233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Poh-Fitzpatrick M. B., Lamola A. A. Direct spectrofluorometry of diluted erythrocytes and plasma: a rapid diagnostic method in primary and secondary porphyrinemias. J Lab Clin Med. 1976 Feb;87(2):362–370. [PubMed] [Google Scholar]
  19. Poh-Fitzpatrick M. B., Piomelli S., Young P., Hsu H., Harber L. C. Rapid quantitative assay for erythrocyte porphyrins. Arch Dermatol. 1974 Aug;110(2):225–230. [PubMed] [Google Scholar]
  20. REDEKER A. G., BRONOW R. S., STERLING R. E. ERYTHROPOIETIC PROTOPORPHYRIA. S Afr J Lab Clin Med. 1963 Dec;14:235–238. [PubMed] [Google Scholar]
  21. Scholnick P., Marver H. S., Schmid R. Erythropoietic protoporphyria: evidence for multiple sites of excess protoporphyrin formation. J Clin Invest. 1971 Jan;50(1):203–207. doi: 10.1172/JCI106474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schwartz S., Johnson J. A., Stephenson B. D., Anderson A. S., Edmondson P. R., Fusaro R. M. Erythropoietic defects in protoporphyria: a study of factors involved in labelling of porphyrins and bile pigments from ALA- 3 H and glycine- 14 C. J Lab Clin Med. 1971 Sep;78(3):411–434. [PubMed] [Google Scholar]
  23. van KAMPEN E., ZIJLSTRA W. G. Standardization of hemoglobinometry. II. The hemiglobincyanide method. Clin Chim Acta. 1961 Jul;6:538–544. doi: 10.1016/0009-8981(61)90145-0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES