title: Photodynamic and light independent action of 8 to 2 carboxylic free porphyrins on some haem-enzymes.
abstract: OBJECTIVE
  skin lesions in cutaneous porphyrias appear to be determined by the structural properties of the porphyrins accumulated. To better understand the relationship between the structure and physicochemical properties of porphyrins and their specific effect on protein configuration, the action of a whole range of 8 to 2 carboxylic porphyrins has been studied. MATERIALS AMD METHODS: delta-aminolevulinic acid dehydratase (ALA-D) and porphobilinogen deaminase (PBG-D) partially purified from bovine liver, were exposed to 10 microM uroporphyrin (Uro), phyriaporphyrin (Phyria), hexaporphyrin (Hexa), pentaporphyrin (Penta), coproporphyrin (Copro) or protoporphyrin (Proto), either in the dark or under UV light. All experiments were performed in the enzyme solutions after removing the porphyrins.
  
  
  RESULTS
  under both illuminating conditions, all porphyrins inactivated the enzymes (20-70% under control values), indicating photodynamic action mediated by oxidative reactions and conformational changes due to direct binding of porphyrins to the protein. Total thiol content in ALA-D was not significantly changed by most porphyrins under UV light, while all porphyrins increase total sulfhydryl groups in PBG-D (23-52% over the control values) indicating changes in the redox status of SH residues. Free amino groups were reduced by all porphyrins in ALA-D (23-56% under controls), instead they were enhanced in PBG-D (23-51% over controls), suggesting protein fragmentation. The formation of molecular aggregates would be the consequence of cross-links between oxidation products, while fragmentation can be attributed to either rupture of disulphur bridges and/or enhancement of free amino groups on the protein enzyme.
  
  
  CONCLUSIONS
  the effect of the porphyrins on enzyme activity, total SH groups and free amino groups content, was different for ALA-D and PBG-D, even under the same illuminating conditions. On the basis of these results, no correlation between enzyme alterations and the physico-chemical properties of porphyrins could be established.
authors.affiliation: Unidad de Porfirias, Hospital Universitario 12 de Octubre, Universidad Complutense de Madrid, Fransisco Gervás 9, 11th A, 28020 Madrid, Spain.
authors.forename: S G
authors.initials: SG
authors.lastname: Afonso
authors.forename: R
authors.initials: R
authors.lastname: Enríquez de Salamanca
authors.forename: A
authors.initials: A
authors.lastname: Batlle
mesh_terms: D000818:Animals; D002417:Cattle; D003306:Coproporphyrins / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000737:chemistry*; D011163:Hydroxymethylbilane Synthase / Q000378:metabolism; D008027:Light; D008099:Liver / Q000201:enzymology; D010100:Oxygen / Q000378:metabolism; D000623:Porphobilinogen Synthase / Q000378:metabolism; D011166:Porphyrins / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014466:Ultraviolet Rays; D014578:Uroporphyrins / Q000494:pharmacology
chemical_list: D003306:Coproporphyrins; D011166:Porphyrins; D013438:Sulfhydryl Compounds; D014578:Uroporphyrins; D006418:Heme; D011163:Hydroxymethylbilane Synthase; D000623:Porphobilinogen Synthase; D010100:Oxygen