Intracellular Energy-Transfer Networks and High-Resolution Respirometry: A Convenient Approach for Studying Their Function.
Marju PuurandKersti TeppAleksandr KlepininLyudmila KlepininaIgor ShevchukTuuli KaambrePublished in: International journal of molecular sciences (2018)
Compartmentalization of high-energy phosphate carriers between intracellular micro-compartments is a phenomenon that ensures efficient energy use. To connect these sites, creatine kinase (CK) and adenylate kinase (AK) energy-transfer networks, which are functionally coupled to oxidative phosphorylation (OXPHOS), could serve as important regulators of cellular energy fluxes. Here, we introduce how selective permeabilization of cellular outer membrane and high-resolution respirometry can be used to study functional coupling between CK or AK pathways and OXPHOS in different cells and tissues. Using the protocols presented here the ability of creatine or adenosine monophosphate to stimulate OXPHOS through CK and AK reactions, respectively, is easily observable and quantifiable. Additionally, functional coupling between hexokinase and mitochondria can be investigated by monitoring the effect of glucose on respiration. Taken together, high-resolution respirometry in combination with permeabilization is a convenient approach for investigating energy-transfer networks in small quantities of cells and tissues in health and in pathology.
Keyphrases
- energy transfer
- protein kinase
- high resolution
- induced apoptosis
- quantum dots
- cell cycle arrest
- gene expression
- healthcare
- reactive oxygen species
- public health
- mass spectrometry
- mental health
- endoplasmic reticulum stress
- cell death
- oxidative stress
- high speed
- skeletal muscle
- transcription factor
- room temperature
- blood glucose
- network analysis