Talk:Zujovic 2019 MitoFit Preprint Arch EA

Comment #1

In Fig1B palmitate group do not seem to be permeabilized even after 6 titrations. There is no significant increase in respiration as it is visible for controls. In Fig2 palmitate group permeabilization is also questionable, because you see an effect on respiration already after 4 ug of digitonin (which is less than in Fig1B) but then there is almost no ADP stimulation: OXPHOS coupling efficiency seems to be less than 50%! And after Rot and Ama the ROX is significantly lower suggesting that endogenous substrates may be also present. Another point in Fig 1A and 2 respiration in OXPHOS for S pathway is smaller than LEAK respiration for N-pathway.

(http://bioblast.at/index.php/OXPHOS_coupling_efficiency

http://wiki.oroboros.at/images/f/fc/Gnaiger_2014_Mitochondr_Physiol_Network_MitoPathways.pdf)

--Sobotka Ondrej (talk) 11:56, 11 June 2019 (CEST)

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Comment #2

What is the reason behind your substrate concentrations (P 8.8 mM, M 4.4mM and S 10 mM)? According to recent work of Sumbalova et. al 2016 it was shown that Malate significantly inhibits S pathway which can be partially overcome by 50 mM succinate. (ref: http://www.bioblast.at/index.php/Sumbalova_2016a_Abstract_MitoFit_Science_Camp_2016) For next experiments you can find SOPs on titration concentrations here:

http://wiki.oroboros.at/images/e/ef/MiPNet09.12_O2k-Titrations.pdf

--Sobotka Ondrej (talk) 11:56, 11 June 2019 (CEST)

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