Porphyrin, chlorin, and bacteriochlorin isothiocyanates: useful reagents for the synthesis of photoactive bioconjugates.
A novel method for conjugating porphyrins and related molecules to proteins has been developed. The method, which involves synthesizing porphyrins, chlorins, and bacteriochlorins bearing a single amine-reactive isothiocyanate group represents a facile system for protein labeling with these photoactive species. Problems associated with the noncovalent binding of porphyrins to proteins are highlighted, and a method for purifying conjugates to yield exclusively covalently bound porphyrin protein species is demonstrated. Biological activity of porphyrin-bovine serum albumin conjugates formed and purified by these methods is demonstrated using laser scanning confocal microscopy.
Sutton
J M
JM
Department of Biological Sciences, University of Essex, Colchester CO4 3SQ, UK.
Clarke
O J
OJ
Fernandez
N
N
Boyle
R W
RW
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D017879:Isothiocyanates / Q000008:administration & dosage / Q000737:chemistry*; D018613:Microscopy, Confocal; D008968:Molecular Conformation; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry; D015394:Molecular Structure; D010777:Photochemistry; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000138:chemical synthesis* / Q000737:chemistry; D011166:Porphyrins / Q000008:administration & dosage / Q000737:chemistry*; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000008:administration & dosage / Q000737:chemistry*
D004337:Drug Carriers; D017879:Isothiocyanates; D015335:Molecular Probes; D017319:Photosensitizing Agents; D011166:Porphyrins; D012710:Serum Albumin, Bovine