Biopolym. Cell. 1995; 11(2):57-68.
http://dx.doi.org/10.7124/bc.0003E1
Supramolecular organisation and activity of smooth muscle myosin light chain kinase
1Filenko A. M., 2Sobieszek A., 1Babiychuk E. B., 1Omelyanuk V. S., 1Danilova V. M., 3Yanovskaya N. B., 1Babenko A. Yu.
  1. Petr Bogach Institute of Physiology
    Taras Shevchenko National University of Kyiv
    2, Academika Glushkova Ave Str., Kyiv, Ukraine, 03187
  2. Institute of Molecular Biology Austrian Academy of Sciences
    Billrothstrasse 11, A-5020 Salzburg, Austria
  3. Palladin Institute of Biochemistry, NAS of Ukraine
    9, Leontovycha Str., Kyiv, Ukraine, 01601

Abstract

The data received by the use of lazer correlation spectroscopy (LCS) suggest that inactive MLCK under conditions close to the physiological exists as mixture of oligomeric and dimeric particles with hydrodynamic diameters about 150 and 20 nm, respectively. These particles should be in dynamic equilibrium. We could not discover noticeable alteration in different kinase species distribution at both kinase or calmodulin excess in solution. At considerable excess of kinase over calmodulin the activity of the former is decreased several times after 5 to 10 min from the moment of its activation. Observed kinase inhibition in the preincubation time (before addition of substrate) must be connected with slow conformational rearrangements of activated kinase molecules which is directly confirmed by the results of our fluorescent studies. Such conformational rearrangements accompanied by the reduction of MLCK towards CaM affinity take place also at equimolar ratios of kinase to CaM, but in this case they are characterised by the lesser depth and insignificant MLCK activity decrease. Slow conformation changes in supramolecular structures of activated kinase are believed to have collective character and to be connected with intensification of mutual influence of kinase molecules leading to reduction of CaM binding.
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