Cellular Uptake Quantification of Metalated Peptide and Peptide Nucleic Acid Bioconjugates by Atomic Absorption Spectroscopy

Kirin, Srećko I.; Ott, Ingo; Gust, Ronald; Mier, Walter; Weyhermüller, Thomas; Metzler‐Nolte, Nils

doi:10.1002/anie.200703994

Cited by 73 publications

(78 citation statements)

References 51 publications

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“…This is rather surprising as the addition of a specific metal complex to a PNA oligomer can be of significant interest. Our group 10 and Krämer et al 11 have, for example, demonstrated that simply attaching a metal complex to PNA bioconjugates can significantly increase the cellular uptake of PNA.…”

Section: Introductionmentioning

confidence: 93%

Synthesis, characterisation and bioimaging of a fluorescent rhenium-containing PNA bioconjugate

et al. 2012

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A new rhenium tricarbonyl complex of a bis(quinoline)-derived ligand (2-azido-N,N-bis((quinolin-2-yl)methyl)ethanamine, L-N(3)), namely [Re(CO)(3)(L-N(3))]Br was synthesized and characterized indepth, including by X-ray crystallography. [Re(CO)(3)(L-N(3))]Br exhibits a strong UV absorbance in the range 300-400 nm with a maximum at 322 nm, and upon photoexcitation, shows two distinct emission bands at about 430 and 560 nm in various solvents (water, ethylene glycol). [Re(CO)(3)(L-N(3))]Br could be conjugated, on a solid phase, to a peptide nucleic acid (PNA) oligomer using the copper(I)-catalyzed azide-alkyne cycloaddition reaction (Cu-AAC, "click" chemistry) and an alkyne-containing PNA building block to give Re-PNA. It was demonstrated that upon hybridisation with a complementary DNA strand (DNA), the position of the maxima and emission intensity for the hybrid Re-PNA·DNA remained mainly unchanged compared to those of the single strand Re-PNA. The rhenium-containing PNA oligomer Re-PNA could be then mediated in living cells where they have been shown to be non-toxic contrary to the general notion that organometallic compounds are usually unstable under physiological conditions and/or cytotoxic. Furthermore, Re-PNA could be detected in living cells using fluorescent microscopy.

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Section: Introductionmentioning

confidence: 93%

Synthesis, characterisation and bioimaging of a fluorescent rhenium-containing PNA bioconjugate

et al. 2012

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“…97 Building on this concept, thymine PNA monomers were modified with dicobalthexacarbonyl derivatives and the preparation of cobalt-containing PNA oligomers is now in The presence of a [Co(bpa)] 2+ derivative into the PNA-NLS bioconjugate ( Figure 12, right, bpa: bispicolylamine) avoids the insertion of a fluorophore. 99 Cobalt is indeed detectable by AAS (atomic absorption spectroscopy), whose background noise in cells is low. Moreover, in contrast to fluorescence microscopy, AAS is a more direct and sensitive detection method, which makes absolute quantification of cellular uptake straight forward.…”

Section: Metal-containing Pnas As Artificial Nucleasesmentioning

confidence: 99%

“…Furthermore, the presence of a metal within a PNA oligomer enables us to quantify its cellular uptake using atomic absorption spectroscopy, as recently demonstrated by our group with a cobalt complex attached to different PNA oligomers. 153 …”

Section: Modulation Of Pna•dna Hybrid Stability Using Metal Complexesmentioning

confidence: 99%

Metal-containing peptide nucleic acid conjugates

2011

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Peptide Nucleic Acids (PNAs) are non-natural DNA/RNA analogues with favourable physicochemical properties and promising applications. Discovered nearly 20 years ago, PNAs have recently re-gained quite a lot of attention. In this Perspective article, we discuss the latest advances on the preparation and utilisation of PNA monomers and oligomers containing metal complexes. These metalconjugates have found applications in various research fields such as in the sequence-specific detection of nucleic acids, in the hydrolysis of nucleic acids and peptides, as radioactive probes or as modulators of PNA˙DNA hybrid stability, and last but not least as probes for molecular and cell biology.

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“…In previous studies, AAS was successfully applied to directly quantify uptake and cellular localization of peptides and peptide nucleic acids (PNAs) bearing cobalt or manganese metal fragments. 15,16 Since, until now, a general comparison of the uptake properties of the branched hCT peptides is lacking. We first labeled the peptides with the fluorophor 7-nitrobenzo-2-oxa-1,3-diazol (NBD) to get some initial insights about their membrane translocation.…”

Section: Introductionmentioning

confidence: 99%

Specific labeling with potent radiolabels alters the uptake of cell‐penetrating peptides

et al. 2009

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Radiolabeled peptides play an important role in radiopharmacy not only as tumor markers but also as transport vectors. Therefore, cell-penetrating peptides (CPP) may serve as very effective delivery tools, as well. Recently, CPP based on the human hormone calcitonin (hCT) have been developed. Especially, branched hCT-peptide sequences turned out to have highly efficient internalization capacities. Labeling these peptides with radionuclides would generate promising new tools for imaging and therapy applications in radiopharmacy. However, the influence of the metal complexation on the internalization capacity of CPP has not been elucidated yet in detail. In this study we quantified the uptake of Ga-DOTA modified hCT-carrier peptides in HeLa cells by using atomic absorption spectroscopy (AAS) and compared the results to the uptake of fluorescently-labeled peptides. Interestingly, we measured different uptake rates depending on the attached label. Unexpectedly, modification with a Ga-DOTA complex can have tremendous effects on the uptake efficiency. The results of these studies support the need of detailed analysis of each carrier peptide/cargo construct, especially in the field of metal complex modified CPP.

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Cellular Uptake Quantification of Metalated Peptide and Peptide Nucleic Acid Bioconjugates by Atomic Absorption Spectroscopy

Cited by 73 publications

References 51 publications

Synthesis, characterisation and bioimaging of a fluorescent rhenium-containing PNA bioconjugate

Synthesis, characterisation and bioimaging of a fluorescent rhenium-containing PNA bioconjugate

Metal-containing peptide nucleic acid conjugates

Specific labeling with potent radiolabels alters the uptake of cell‐penetrating peptides

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