Аннотация:In [1] it has been shown that under hypotonic condition (medium osmolarity 120
mosM) at room temperatures (15-22 °C) osmotic pressure rearranges mitochondrial
OxPhos system and makes it operate as complete supercomplex, including electron-
transport chain proteins and ATP-syntetase complex. A purpose of present work
was to investigate whether such conditions give OxPhos system any advantages, for
example some energy gain. Measuring parameter ADP/O (which shows an efficiency
of phosphorylation) in wide temperature range, we found two temperature regions
where ADP/O is maximal and close to theoretical value. (Fig.1) One of them is the
range of physiological temperatures 30-40 °C.
Unexpectedly another range proved to be very narrow (not wider than 1 °C)
vicinity of 19 °C. We found out that ADP/O at 19 °C rises abruptly up to the
theoretical level of 2, while at neighbouring temperatures ADP/O is about 1,5.
General reduction of ADP/O level at room temperatures is shown to be caused by
uncoupling effect of endogenous fatty acids. They are endogenous substrates of
mitochondrial oxidation. To reproduce physiological conditions, we deliberately have
not removed them from membranes during mitochondrial isolation.
The special temperature 19 °C is characterized by stronger immunity of
OxPhos system to uncoupling effect of fatty acids. If fatty acids are removed from
membranes, the system is able to operate with maximal efficiency at all room
temperatures. That immunity is so strong that even when exogenous fatty acids
are added up to almost complete uncoupling, ADP/O at 19 °C in comparison with
neighboring temperatures is steel significantly rised.
The special temperature observed suddenly coincides with narrow temperature
gap where temperature-dependent structural rearrangement of mitochondrial
membrane has been detected in [2] under the same conditions. Stronger aggregation
of membrane proteins has been found at 19±2 °C by measuring tryptophan
fluorescence quenching by pyrene localised in lipid membrane component. (Fig.2)
Thus, near 19 °C OxPhos system operating as supercomplex displays structural
and functional special features. It is protected (partially or fully) from mild
uncoupling.
The anomalous narrowness of temperature gap where both effects are observed
is of great question. Special state of OxPhos system under conditions of osmotic
pressure in immediate vicinity of 19 °C could be regarded as intermediate state of
supercomplex as highly organized structure. It is worth to notice that 19 °C is also
the special temperature for water (e.g. there is OH-bond energy anomaly at this
temperature [3]). So one could link the effects discussed with melting of highly
organized proteins hydration shell.