Spectroscopic studies on the interaction of a water soluble porphyrin and two drug carrier proteins.
The interaction of meso-tetrakis(p-sulfonatophenyl)porphyrin (TSPP) sodium salt to human serum albumin and beta-lactoglobulin was studied by steady-state and dynamic fluorescence at different pH of aqueous solutions. The formation of TSPP J-aggregates and a noncovalent TSPP-protein complex was monitored by fluorescence titrations, which depend on pH and on the protein nature and concentration. The complex between TSPP and protein displays a heterogeneous equilibrium with large changes in the binding strength versus pH. The large reduction of the effective binding constant from pH 2 to 7 suggests that electrostatic interactions are a major contribution to the binding of TSPP to the aforementioned proteins. TSPP aggregates and TSPP-protein complex exhibit circular dichroism induced by the presence of the protein. Circular dichroism spectra in the ultraviolet region show that the secondary structure of both proteins is not extensively affected by the TSPP presence. Protein-TSPP interaction was also examined by following the intrinsic fluorescence of the tryptophan residues of the proteins. Fluorescence quenching by acrylamide and TSPP itself also point to small changes on the protein tertiary structure and a critical distance R(0) approximately 56 A, between tryptophan and bound porphyrin, was estimated using the long distance Förster-type energy transfer formalism.
Andrade
Suzana M
SM
Centro de Química Estrutural, Complexo 1, Instituto Superior Técnico, 1049-001 Lisboa, Portugal. sandrade@popsrv.ist.utl.pt
Costa
Sílvia M B
SM
D000178:Acrylamides / Q000737:chemistry; D002942:Circular Dichroism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry; D008956:Models, Chemical; D011166:Porphyrins / Q000737:chemistry*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012709:Serum Albumin / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry / Q000379:methods*; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry; D014466:Ultraviolet Rays; D014867:Water / Q000737:chemistry*
D000178:Acrylamides; D007782:Lactoglobulins; D011166:Porphyrins; D012709:Serum Albumin; D014867:Water; C016136:tetraphenylporphine sulfonate; D014364:Tryptophan