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Oligo- and polypeptide conjugates of cationic porphyrins: binding, cellular uptake, and cellular localization

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Abstract

Recently, we have characterized the DNA and nucleoprotein (NP) binding of bis(4-N-methylpyridyl)-15,20-di(4-carboxyphenyl)porphyrin (BMPCP) and meso-tri(4-N-methylpyridyl)-mono(4-carboxyphenyl)porphyrin (TMPCP) and their tetrapeptide conjugates (BMPCP-4P2 and TMPCP-4P, respectively). In this work, we investigated the interaction of TMPCP conjugated to the tetrapeptide branches of branched chain polymeric polypeptide with poly-l-lysine backbone (AK) with DNA or NP using spectroscopic methods. Analysis of absorption spectra revealed the external binding but no intercalation of TMPCP-AK to DNA. There was no evidence for the interaction between TMPCP-AK and encapsidated DNA. Furthermore, we examined the cellular uptake of BMPCP and TMPCP and their tetra- or polypeptide conjugates by flow cytometry and analyzed how charge, size, and structure of the compounds affect their incorporation. In comparison, liposomal association constants of these derivatives were determined. BMPCP-4P2 accumulated the most, and porphyrins with two positive charges (BMPCP and BMPCP-4P2) showed better accumulation than the tri-cationic TMPCP or TMPCP-4P. Cellular uptake of polycationic TMPCP-AK was significantly lower than that of the free or tetrapeptide conjugated derivatives. The subcellular localization of all the five compounds was investigated in co-localization studies by confocal microscopy with special attention to their nuclear localization. Neither free nor conjugated BMPCP or TMPCP was co-localized with nuclear marker. Instead, these derivatives showed co-localization with lysosomal and mitochondrial fluorescent probes. TMPCP-AK conjugate had different localization patterns appearing mainly in mitochondria and cytoplasmic vesicles. Our results may contribute to the further design of DNA-targeting porphyrin-based constructs.

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Acknowledgements

This work was supported by research grant Országos Tudományos Kutatási Alapprogramok (OTKA) K104045 and K104275.

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Authors and Affiliations

  1. Institute of Biophysics and Radiation Biology, Semmelweis University, POB 263, Budapest, 1444, Hungary

    Ádám Orosz, Levente Herényi & Gabriella Csík

  2. Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Eötvös Loránd University, Pázmány Péter sétány 1/A, Budapest, 1117, Hungary

    Szilvai Bősze, Gábor Mező & Ildikó Szabó

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  1. Ádám Orosz
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  2. Szilvai Bősze
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  3. Gábor Mező
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  4. Ildikó Szabó
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  5. Levente Herényi
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  6. Gabriella Csík
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Correspondence to Gabriella Csík.

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Orosz, Á., Bősze, S., Mező, G. et al. Oligo- and polypeptide conjugates of cationic porphyrins: binding, cellular uptake, and cellular localization. Amino Acids 49, 1263–1276 (2017). https://doi.org/10.1007/s00726-017-2428-z

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  • DOI: https://doi.org/10.1007/s00726-017-2428-z

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