Tag Archives: recombinant protein

Influence of human HB-EGF secreted form on cells with different EGFR and ErbB4 quantity

O. I. Krynina, N. V. Korotkevych, A. J. Labyntsev,
S. I. Romaniuk, D. V. Kolybo, S. V. Komisarenko

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

Received: 18 July 2019; Accepted: 13 August 2019

HB-EGF is one of the most potent ligands of EGFR and ErbB4 receptors. This growth factor plays a pivotal role in many cellular processes, but its effect differs from one cell type to another and remains not fully understood. The aim of this work was to investigate the dependence between the rate of HB-EGF mediated cell proliferation and activation of EGFR and ErbB4 receptors. Therefore, the effects of human recombinant sHB-EGF (rsHB-EGF) on the proliferation of cell lines with different EGFR and ErbB4 quantity and ratio, as well as activation of the MARK-cascade p38 and ERK1/2 (p42/44) kinases, were analyzed. For comparison, a similar study of the effect of native sHB-EGF secreted by human histiocytic lymphoma cells U937 during co-cultivation with different cell lines was performed.
It was proved that cell proliferation in response to sHB-EGF depends not only on the quantity but also on the ratio of EGFR and ErbB4. It was shown that signaling through ErbB4 is associated with activation of p38 kinase and signaling through EGFR associated with activation of ERK1/2 (p42/44) kinase. We assume the existence of two different mechanisms for sHB-EGF-mediated stimulation of cell proliferation, and the simultaneous launch of these mechanisms provides a maximal proliferative response. The results of this study support the feasibility of creating anti-proliferative drugs that target ErbB4.

Specificity and sensitivity of the new test for serological evaluation of tuberculosis using MPT83-MPT63 fusion antigen

A. A. Siromolot1,2, T. O. Chudina2, I. S. Danilova3,
O. M. Rekalova4, D. V. Kolybo1,2, S. V. Komisarenko2

1ESC Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3NSC Institute of Experimental and Clinical Veterinary Medicine, Kharkiv, Ukraine;
4SI National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky National Academy of Medical Sciences of Ukraine, Kyiv;
e-mail: saa0205@ukr.net

The aim of this work was to characterize the experimental test-system based on MPT83-MPT63 fusion antigen with reference commercial diagnostics and investigate the basic characteristics of enzyme-linked immunosorbent assay (ELISA) tests such as specificity and sensitivity. In addition, we investigated the correlation of biochemical and immunological parameters of blood samples of patients with tuberculosis, which was correctly diagnosed by those test-systems. It was shown that the developed test-system match the existing ones regarding the criteria of reliability and the basic requirements of ELISA kits. Recommendations for the testing of serum samples and indications for the use of the proposed serological diagnostic methods has been suggested.

Investigation of tomato (Solanum lycopersicum) aminotransferases involved in biosynthesis of branched-chain-amino-acids

A. S. Kochevenko1,2, A. R. Fernie2

1Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv;
2Max Planck Institute of Molecular Plant Physiology, Golm, Germany;
e-mail: andkochevenko@gmail.com

This study presents evidence for the role of ВСАТ3 and ВСАТ4 proteins in the synthesis of branched-chain-amino-acids in tomato Solanum lycopersicum. ВСАТ3 and ВСАТ4 genes were located­ on tomato chromosomal map by RFLP method (restriction fragment length polymorphism). Using confocal microscopy it was shown that BCAT3-GFP and BCAT4-GFP fusion proteins were localised in chloroplasts. It was observed that these aminotransferase isoforms exhibited distinct kinetic­ properties and a differential expression pattern of mRNA levels in various tomato tissues.