a brief introduction to pka ph fluorescent probe
TRANSCRIPT
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A brief introduction to pKa
& pH Fluorescent Probe
汇报人:Yu mengyi
指导人:Prof. Zhao
Dr. Hong mei
2014/9/9
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pKa • Acidities are quantified by pKa values. The pKa
of an acid HX is defined as
• The larger the pKa, the less acidic the compound.
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Factors
• Medium Effects
• Hybridization Effects
• Substituent Effects
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Medium Effects
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Hybridization Effects
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Substituent Effects
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Measurement
• Potentiometric titration
• Conductometry
• Spectrophotometry
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Mitochondria-Immobilized pH-Sensitive Off−On Fluorescent Probe
Min Hee Lee, Nayoung Park, Chunsik Yi, Ji Hye Han, Ji Hye Hong, Kwang Pyo Kim,Dong Hoon Kang, Jonathan L. Sessler, Chulhun Kang, and Jong Seung Kim
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Background
Youle, R. J.; Narendra, D. P. Mol. Cell. Biol. 2011, 12, 9−14. Kim, I.; Rodriguez-Enriquez, S.; Lemasters, J. J. Arch. Biochem.Biophys. 2007, 462, 245−253.
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pH sensing mechanism
1
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In vitro
2 1
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Stability and MTT
Na+, K+, Ca2+, Zn2+, Mg2+, Mn2+, Cu2+, Fe2+, Fe3+, thiols (GSH, Cys, Hcy)
MTT assay
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in vivo--selectivity
MitoTracker Red(MTR)
mitochondrion-specific
fluorescent probe
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Immobilization
CCCP induces an uncoupling of the mitochondrial membrane potential (Δψm)
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Real-time monitoring LysoTracker Red(LTR) Pepstatin A is a protease inhibitor and was used to delay mitochondrial degradation by proteases in the autolysosomes.
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Summary • The probe allow the direct and reliable
mitochondrial pH measurement in whole cells.
• Achieve the real time monitoring of pH changes associated with the mitochondrial acidification and fusion.
• Act a means of distinguishing between physiological and pathological states or screening potential new mitochondria-targeting drugs.
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Thank you !