Tag Archives: fibrin polymerization

Men of the molecules

In memoriam of Prof. Russell Doolittle,
Prof. Eduard Lugovskoi and their friendship that outlive both of them

V. O. Chernyshenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: bio.cherv@gmail.com

Received: 02 July 2020; Accepted: 21 July 2020

In memoriam Eduard Lugovskoi and Russell Doolittle we are referring to several episodes of their life and work.  Russell Doolittle an American biochemist and his friend and colleague Ukrainian scientist Eduard Lugovskoi, both studied fibrinogen structure and functions and finally united their efforts in the revealing of the new mechanism of intramolecular interactions of fibrin molecule through coiled-coil region. The results of their common work and discussions were included to the article “The fibrin Bβ125-135 site is involved in the lateral association of protofibrils”. Valuable part of the communication dedicated to the poetry of Eduard Lugovskoi that inspired both of scientists in work and life. We are providing some remembrance of their collaboration, their letters sent to each other, fragments of handwriting and common photo of Russell Doolittle and Eduard Lugovskoi.

Calixarene methylene bisphosphonic acids as promising effectors of biochemical processes

S. V. Komisarenko1, S. O. Kosterin1, E. V. Lugovskoy1, V. I. Kalchenko2

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: kinet@biochem.kiev.ua;
2Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: vik@ioch.kiev.ua

This interdisciplinary study, performed with participation of research workers of Palladin Institute of Biochemistry and Institute of Organic Chemist­ry of NAS of Ukraine, is devoted to analysis of biochemical effects of some calixarene methylene bisphosphonic acids (cyclic phenol oligomers) on two well-known biological phenomenons – Mg2+-dependent ATP hydrolysis (myosin subfragment-1 of myometrium smooth muscle was used as an example) and fibrin polymerization.
Calix[4]arene С-97 (calix[4]arene methylene bisphosphonic acids) is a macrocyclic substance, which contains intramolecular highly ordered lipophilic cavity formed by four aromatic rings, one of which is functionalized at the upper rim with methylene bisphosphonic group. At concentration of 100 µM, this substance was shown to effectively inhibit ATPase activity of pig myometrium myosin subfragment-1 (inhibition coefficient І0.5 = 83 ± 7 µM). At the same time, this calix[4]arene causes significant (vs. control) increase of myosin subfragment-1 hydrodynamic diameter, which may indicate formation of an intermolecular complex between calixa­rene and myosin head. Computer simulation methods (docking and molecular dynamics with addition of grid technologies) enabled to elucidate the grounds of intermolecular interactions between calix[4]arene С-97 and myometrium myosin subfragment-1, that involve hydrophobic, electrostatic and π-π-stacking interactions, some of which are close to the ATPase active centre. In view of the ability of calixarenes to penetrate into the cell and their low toxicity, the results obtained may be used as a basis for further development of a new generation of supramolecular effectors (starting from the above mentioned substances, in particular calix[4]arene С-97) for regulation of smooth muscle contractile activity at the level of ATP dependent actin-myosin interaction.
Calix[4]arenes bearing two or four methylenebisphosphonic acid groups at the macrocyclic upper rim have been studied with respect to their effects on fibrin polymerization. The most potent inhibitor proved to be calix[4]arene tetrakis-methylene-bis-phosphonic acid (C-192), in which case the maximum rate of fibrin polymerization in the fibrinogen + thrombin reaction decreased by 50% at concentrations of 0.52·10-6 M (IC50). At this concentration, the molar ratio of the compound to fibrinogen was 1.7 : 1. For the case of desAB fibrin polymerization, the IC50 was 1.26·10-6 M at a molar ratio of C-192 to fibrin monomer of 4 : 1. Dipropoxycalix[4]-arene bis-methylene-bis-phosphonic acid (C-98) inhibited fibrin desAB polymerization with an IC50 = 1.31·10-4 M. We hypothesized that C-192 blocks fibrin formation by combining with polymerization site ‘A’ (Aa17–19), which ordinarily initiates protofibril formation in a ‘knob-hole’ manner. This suggestion was confirmed by an HPLC assay, which showed a host–guest inclusion complex of C-192 with the synthetic peptide Gly-Pro-Arg-Pro, an analogue of site ‘A’. Further confirmation that the inhibitor was acting at the initial step of the reaction was obtained by electron microscopy, with no evidence of protofibril formation being evident. Calixarene C-192 also doubled both the prothrombin time and the activated partial thromboplastin time in normal human blood plasma at concentrations of 7.13·10-5 and 1.10·10-5 M, respectively. These experiments demonstrate that C-192 is a specific inhibitor of fibrin polymerization and blood coagulation and can be used for the design of a new class of antithrombotic agents.

Laureates of the Palladin Prize of the National Academy of Sciences of Ukraine (2001, 2003 years)

V. M. Danilova, R. P. Vynogradova

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: valdan@biochem.kiev.ua

In 2001 L. L. Gromashevska (1922-2009), Head of Laboratory at L. V. Gromashevsky Institute of Epidemiology and Infectious Diseases, NAMS of Ukraine and A. S. Mikosha, Head of Laboratory at V. P. Komisarenko Institute of Endocrinology and Metabolism, NAMS of Ukraine were awarded the Palladin Prize of NAS of Ukraine for the series of works “The regulation of biochemical processes in norm and pathology“, which presents the results of the research on hormone and enzyme biochemistry, signaling pathways in cells under different physiological and pathological conditions.
In 2003 S. V. Komisarenko, E. V. Lougovskoy and I. M. Kolesnikov, Director and staff of Palladin Institute of Biochemistry, NAS of Ukraine, respectively, were awarded the Palladin Prize of NAS of Ukraine for the series of works “Immunochemical analysis of mechanisms of fibrin polymerization and fibrinolysis“. The paper presents an analysis of scientific activity and laureates’ biographical information.