title: Further evidence on the photodynamic and the novel non-photodynamic inactivation of uroporphyrinogen decarboxylase by uroporphyrin I.
abstract: The action of uroporphyrin I (URO I) on the activity of red cell uroporphyrinogen decarboxylase (URO-D) in the dark and under UV light was studied. Light-dependent-and light-independent inactivation was observed. Both effects increased at increasing concentrations of URO I, the former reached its maximum at 150 microM of sensitizer. At 100 microM of URO I, both light and dark inactivation were temperature dependent amounting to about 50% at 30-37 degrees C. The velocity of dark inactivation increased with increasing temperature in the range of 0 to 45 degrees C. Photoinactivation can be ascribed to primary oxidation of essential amino acids, very likely histidyl residues, followed by secondary inter or intrapeptide cross-linking. Dark inactivation could be the result of both oxidation and cross-linking (although to a less degree than that produced by light) and also direct inhibition of the enzyme by induced conformational changes at its active site through binding of the porphyrin to the protein. When the action of URO I was tested on partially purified URO-D, the enzyme appeared to be more susceptible to the dark than to the light effect.
authors.affiliation: Centro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), University of Buenos Aires, Argentine.
authors.forename: S G
authors.initials: SG
authors.lastname: Afonso
authors.forename: S
authors.initials: S
authors.lastname: Chinarro
authors.forename: R E
authors.initials: RE
authors.lastname: de Salamanca
authors.forename: A M
authors.initials: AM
authors.lastname: Batlle
mesh_terms: D003624:Darkness; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D017319:Photosensitizing Agents / Q000494:pharmacology*; D014466:Ultraviolet Rays; D014575:Uroporphyrinogen Decarboxylase / Q000037:antagonists & inhibitors* / Q000097:blood / Q000528:radiation effects; D014578:Uroporphyrins / Q000494:pharmacology*
chemical_list: D017319:Photosensitizing Agents; D014578:Uroporphyrins; C034103:uroporphyrin I; D014575:Uroporphyrinogen Decarboxylase