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Bioorganic chemistry: Institutes, journals, publications, a short scientific metric analysis on the 50th anniversary of the Institute of Natural Compound Chemistry, Institute of Bioorganic Chemistry

Russian Journal of Bioorganic Chemistry : Volume 35, Issue 1, January 2009

Russian Journal of Bioorganic Chemistry : Volume 34

Russian Journal of Bioorganic Chemistry : Volume 33

Russian Journal of Bioorganic Chemistry : Volume 32

Russian Journal of Bioorganic Chemistry : Volume 31

Russian Journal of Bioorganic Chemistry : Volume 30

Russian Journal of Bioorganic Chemistry : Volume 29

Russian Journal of Bioorganic Chemistry : Volume 28

Russian Journal of Bioorganic Chemistry : Volume 27

Russian Journal of Bioorganic Chemistry : Volume 26

The biotin-thyroxin conjugate as a bifunctional ligand of binding proteins

Content Provider	Springer Nature Link
Author	Navakouski, M. J. Vashkevich, I. I. Sviridov, O. V.
Copyright Year	2009
Abstract	The conjugate of the residue of vitamin H (biotin, Bt) with the hormone of thyroid gland thyroxin (T$_{4}$) was prepared by N-acylation of N-(3-aminopropyl)biotin amide with N-hydroxysuccinimide ester of N-acetyl thyroxin. The interactions of the Bt-T$_{4}$ conjugate with one or simultaneously with two binding proteins with affinity to Bt or T$_{4}$ in solution and on a solid phase were studied by electron spectroscopy, enzyme immunoassay, and computer modeling. Bt-T$_{4}$ was specifically fixed in the Bt-binding site of the streptavidin molecule via a large number of hydrogen bonds and hydrophobic interactions. The maximum of the streptavidin fluorescence shifted to a long-wave area and its intensity decreased as a result of complex formation. The degree of quenching of the protein emission was significantly higher than that of the streptavidin-Bt complex. Additional fluorescence quenching resulted from interactions which were sensitive to pH, ionic strength, and detergents and stabilized the position of the thyroxin part of the conjugate near Trp120 of streptavidin in its complex with Bt-T$_{4}$. The Bt-T$_{4}$ conjugate also formed a specific equimolar complex with T$_{4}$-binding human globulin (TBG) by the same mechanism as that for T$_{4}$. The Bt residue did not participate in the interactions which changed characteristics of the TBG fluorophores. The Bt-T$_{4}$ conjugate was bound to avidin on a solid phase in the solid phase enzyme immunoassay owing to its biotin function, whereas its thyroxin part was exposed to a solution and interacted with polyclonal antibodies to T$_{4}$. The intact T$_{4}$ competitively inhibited this interaction after its addition to the system. Bt-T$_{4}$ also exhibited its bifunctional activity in other immune analytic system. The conjugate bound streptavidin was labeled with Eu$^{3+}$-chelate and subsequently formed a three component complex with participation of a monoclonal antibody to T$_{4}$ immobilized on a solid phase. Free T$_{4}$ inhibited the thyroxin function of the conjugate bound to the labeled streptavidin proportionally to its concentration in a sample of human blood serum. Parameters of the immunofluorescent analysis demonstrated that the streptavidin-Bt-T$_{4}$ complex was actively bound to the T$_{4}$-antibody, but had practically no interaction with serum T$_{4}$-binding proteins, including TBG. Probably, nonspecific interactions of the T$_{4}$ residue with streptavidin in its complex with Bt-T$_{4}$, along with steric factors, complicated penetration of thyroxin in this complex into active sites of TBG and other T$_{4}$-binding proteins of blood serum. The Bt-T$_{4}$ stable conjugate was synthesized according to a plain scheme and could be used as a bifunctional ligand of binding proteins in biochemical studies and immune analytical systems for medicinal diagnostics.
Starting Page	163
Ending Page	175
Page Count	13
File Format	PDF
ISSN	10681620
Journal	Russian Journal of Bioorganic Chemistry
Volume Number	35
Issue Number	2
e-ISSN	1608330X
Language	English
Publisher	SP MAIK Nauka/Interperiodica
Publisher Date	2009-03-31
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	biotin-thyroxin conjugate streptavidin bifunctional ligand fluorescence quenching lanthanide immunofluorimetric analysis Biomedicine general Organic Chemistry Bioorganic Chemistry Biochemistry
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Biochemistry

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