Effects of site specific amino acid exchanges in hirudin on the thrombin-hirudin interaction.
For the identification of the primary binding site of hirudin for thrombin we generated hirudin mutants with site directed amino acid substitutions with the help of recombinant DNA technology. Preliminary results indicate, that lys (47) may be directly involved in the hirudin-thrombin interaction: 1. The mutant glu (47) shows a Ki-value which is increased by two orders of magnitude (1.6.10(-9) M); 2. Incubation of mutant ala(48) with endoproteinase lys-C results in proteolysis of the newly formed peptide bond lys(47)-ala(48), whereas all other peptide bonds (lys-X) are not accessible.
Köhler
S
S
Institute of Organic Chemistry and Biochemistry, Darmstadt Technical College, FRG.
Dodt
J
J
D000596:Amino Acids / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004347:Drug Interactions; D010450:Endopeptidases; D005796:Genes; D006629:Hirudins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D013379:Substrate Specificity; D013917:Thrombin / Q000037:antagonists & inhibitors*
D000596:Amino Acids; D006629:Hirudins; D015345:Oligonucleotide Probes; D010450:Endopeptidases; D013917:Thrombin