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Volume 272, Number 33, Issue of August 15, 1997 pp. 20328-20331
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

COMMUNICATION:
Nuclear Targeting of Chlorin e6 Enhances Its Photosensitizing Activity

(Received for publication, April 15, 1997, and in revised form, June 13, 1997)

Tamara V. Akhlynina Dagger , David A. Jans § , Andrey A. Rosenkranz Dagger , Natalya V. Statsyuk , Irina Y. Balashova , Gabor Toth par , Imre Pavo par , Andrey B. Rubin and Alexander S. Sobolev Dagger

From the Dagger  Biophysical Laboratory, Russian Institute of Agricultural Biotechnology, Timiryazevskaya Street 42, 127550, Moscow, Russia, the § John Curtin School of Medical Research, Division for Biochemistry and Molecular Biology, Nuclear Signaling Laboratory, Canberra City, A.C.T. 2601, Australia, the  Department of Biophysics, Biological Faculty, Moscow State University, 119899 Moscow, Russia, and the par  Institute for Medical Chemistry, Szeged Medical University, Szeged, H-6720 Hungary

Although photosensitizers, molecules that produce active oxygen species upon activation by visible light, are being extensively used in photodynamic therapy to treat cancer and other clinical conditions, problems include normal cell and tissue damage and associated side effects, which are attributable in part to the fact that cytotoxic effects are largely restricted to the plasma membrane. We have previously shown that the photosensitizer chlorin e6 has significantly higher photosensitizing activity when present in conjugates containing specific ligands and thus able to be internalized by receptor-expressing cells. In this study we use insulin-containing conjugates to which variants of the simian virus SV40 large tumor antigen nuclear localization signal (NLS) were linked to target chlorin e6 to the nucleus, a hypersensitive site for active oxygen species-induced damage. NLSs were either included as peptides cross-linked to the carrier bovine serum albumin or encoded within the sequence of a beta -galactosidase fusion protein carrier. The results for photosensitization demonstrate clearly for the first time that NLSs increase the photosensitizing activity of chlorin e6, maximally reducing the EC50 by a factor of over 2000-fold. This has wide-reaching implications for achieving efficient cell type-specific photodynamic therapy.


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