Biopolym. Cell. 1992; 8(4):20-30.
Structure and Function of Biopolymers
Dipeptides conteining m- and p-fluorophenylalanine. Synthesis separation on optical isomers and study of their substrate properties towards thrombin
1Poyarkova S. A., 1Kukhar V. P., 1Kolicheva M. T., 1Hrapunov S. N., 1Dragan A. I.
  1. Institute of Bioorganic Chemistry and Petrochemistry, NAS of Ukraine
    1, Murmans'ka Str., Kyiv, Ukraine, 02094


In order to search of new substrates and inhibitors for thrombin DL-and LL-stereoisomers of dipeptide Tos-Phe-(X)-Arg-OCH3 (where X-tn- or p-fluorophenylalanine) were synthesized by classical methods of peptide chemistry from racemate tosylfluorophenylalanine. Tos-L-Phe(pF)-Arg-OCH3 showed substrate's activation at concentration [S] >KM and inhibition by substrate at [S] < KM Tos-L-Phe(mF)-Arg-OCH3 has no bioregulatory effect. Isomers containing D-fluorophenylalanine are not split by thrombin. Study of absorption spectrum and difference in spectrums of fluorescence showed different conformational flexibility of DL and LL isomers. Bathochromic shift in absorption spectrum and fluorescent spectrums of m-fluorophenylalanine is differ from the same of dipeptide, containing p-fluorophenylalanine. The connection between conformational flexibility and substrate's properties of dipeptides is discussed.


[1] Fenton JW, Landis BH, Walz DA et al. Human thrombin: Preparative, evolution, structural properties, and enzymic specificity. Chem. and physiol. of human plasma proteins. Ed. D. H. Bing. New York : Pergamon press, 1979: 151-73.
[2] Fenton JW 2nd. Thrombin specificity. Ann N Y Acad Sci. 1981;370:468-95.
[3] Liem RKH, Scheraga HA. Mechanism of action of thrombin on fibrinogen. III. Partial mapping of the active sites of thrombin and trypsin. Arch Biochem Biophys. 1973;158(1):387–95.
[4] Meinwald YC, Martinelli RA, van Nispen JW, Scheraga HA. Mechanism of action of thrombin on fibrinogen. Size of the A alpha fibrinogen-like peptide that contacts the active site of thrombin. Biochemistry. 1980;19(16):3820-5.
[5] Liem RKH, Scheraga HA. Mechanism of action of thrombin on fibrinogen. IV. Further mapping of the active site of thrombin and trypsin. Arch Biochem Biophys 1974;160(1):333–9.
[6] Pozsgay M, Szab? G, Bajusz S, Simonsson R, G?sp?r R, El?di P. Study of the specificity of thrombin with tripeptidyl-p-nitroanilide substrates. Eur J Biochem. 1981;115(3):491-5.
[7] Bajusz S, Barab?s E, Tolnay P, Sz?ll E, Bagdy D. Inhibition of thrombin and trypsin by tripeptide aldehydes. Int J Pept Protein Res. 1978;12(4):217-21.
[8] Fenton JW 2nd. Regulation of thrombin generation and functions. Semin Thromb Hemost. 1988;14(3):234-40.
[9] Poiarkova SA, Kibirev VK, Serebriany? SB. Inhibitory effect of methyl esters of arginine-containing oligopeptides on thrombin and trypsin. Ukr Biokhim Zh. 1987;59(5):5-11.
[10] Poiarkova SA, Kibirev NK, Serebriany? SB. Inhibition of the proteolytic activity of thrombin by methyl esters of arginine-containing oligopeptides. Ukr Biokhim Zh. 1986;58(6):3-8.
[11] Klesov AA, Berezin IV. Use of integrated rate equations for the determination of the kinetic constants of enzyme reactions. Biokhimiia. 1972;37(1):170-83.
[12] Bode W, Mayr I, Baumann U, Huber R, Stone SR, Hofsteenge J. The refined 1.9 A crystal structure of human alpha-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J. 1989;8(11):3467-75.
[13] Bakhshiev NG. Spectroscopy of intermolecular interactions. Leningrad: Nauka, 1972. 265 p.
[14] Sverdlov OV. Electronic spectra in organic chemistry. Khimiya, 1973. 248 p.
[15] Lakowicz JR, Principles of Fluorescence Spectroscopy. Plenum Press, New York, London, 1983.
[16] Strukova SM, Semenova OA, Kireeva EG. Regulation of alpha and beta/gamma-thrombin activity by heparin and indole. Biokhimiia. 1980;45(4):738-46.