TAILIEUCHUNG - Báo cáo khoa học: Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor

Research on cell-penetrating peptides for the intracellular delivery of por-phyrin compounds has mainly focused on the use of trans-activator of transcription (TAT)-derived peptides and, to a lesser extent, on proline-rich peptides and phosphorescent metalloporphyrins. In this article, we describe a novel phosphorescent oxygen-sensitive probe for intracellular use which comprises a bactenecin 7 peptide fragment (15–24) conjugated with the uncharged monofunctional derivative of Pt(II) coproporphyrin I (PEPP0) | ỊFEBS Journal Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor Ruslan I. Dmitriev1 Honorata M. Ropiak1 Dmitri V. Yashunsky2 Gelii V. Ponomarev2 Alexander V. Zhdanov1 and Dmitri B. Papkovsky1 1 Biochemistry Department University College Cork Ireland 2 Institute of BiomedicalChemistry Russian Academy of MedicalSciences Moscow Russia Keywords bactenecin cell-penetrating peptide intracellular oxygen porphyrin respirometry Correspondence R. I. Dmitriev D. B. Papkovsky Biochemistry Department University College Cork Cavanagh Pharmacy Building Cork Ireland Fax 353 21 4901698 353 21 4901698 Tel 353 21 4901699 353 21 4901698 E-mail Received 21 July 2010 revised 1 September 2010 accepted 7 September 2010 doi Research on cell-penetrating peptides for the intracellular delivery of porphyrin compounds has mainly focused on the use of trans-activator of transcription TAT -derived peptides and to a lesser extent on proline-rich peptides and phosphorescent metalloporphyrins. In this article we describe a novel phosphorescent oxygen-sensitive probe for intracellular use which comprises a bactenecin 7 peptide fragment 15-24 conjugated with the uncharged monofunctional derivative of Pt II coproporphyrin I PEPP0 . This probe provides efficient loading of various mammalian cells including PC12 HCT116 SH-SY5Y and HeLa via cell-type-dependent uptake mechanisms. The conjugate displays a similar distribution in cytoplasm and mitochondria which allows local oxygen levels to be monitored. Respiratory responses of PC12 cells loaded with the conjugate measured on a time-resolved fluorescent reader showed significant cell deoxygenation in response to uncoupling by carbonyl cyanide 4- trifluoromethoxy phen-ylhydrazone and external hypoxia. Treatment with mitochondrial inhibitors led to a decrease in cell deoxygenation. Although the biophysical properties of this conjugate are

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