Intracellular Energy-Transfer Networks and High-Resolution Respirometry: A Convenient Approach for Studying Their Function

International Journal of Molecular Sciences

Aleksandr Klepinin¹  Lyudmila Klepinina¹  Kersti Tepp¹  Marju Puurand¹  Tuuli Kaambre¹  Igor Shevchuk¹ 

[1] Laboratory of Chemical Biology, National Institute of Chemical Physics and Biophysics, Akadeemia tee 23, 12618 Tallinn, Estonia;

关键词: mitochondria;    energy-transfer networks;    creatine kinase;    adenylate kinase;    hexokinase;    oxidative phosphorylation;    high-resolution respirometry;   

DOI  :  10.3390/ijms19102933

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown