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Primary structures of new 'iso-hirudins'
M Scharf1, J Engels, D Tripier
1Institut für organische Chemie, Johann Wolfgang v. Goethe Universität, Frankfurt, FRG.
FEBS Letters
|September 11, 1989
Summary
Researchers isolated and sequenced novel isohirudins from leech crude hirudin using HPLC and fragmentation techniques. These compounds exhibit comparable thrombin inhibition activity, raising questions about their genetic origin.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Crude hirudin, a polypeptide mixture from leeches, is a known anticoagulant.
- Understanding the diversity and origin of hirudin variants is crucial for drug development.
Purpose of the Study:
- To separate and determine the primary sequences of novel isohirudins from crude hirudin.
- To assess the biological activity of these isohirudins.
- To investigate the evolutionary origin of isohirudins.
Main Methods:
- Microbore High-Performance Liquid Chromatography (HPLC) for separation.
- Amino acid analysis and N-terminal microsequencing for primary structure determination.
- Chemical and enzymatic fragmentation for sequence elucidation.
- Thrombin inhibition assays for biological activity assessment.
Main Results:
- Successfully separated and identified three new isohirudins (Ia-IIIb') from crude hirudin.
- Determined the complete primary amino acid sequences of isohirudins Ia-IIIb'.
- All isolated isohirudins demonstrated comparable biological activity in inhibiting thrombin.
Conclusions:
- The study provides the primary sequences for novel isohirudins.
- Isohirudins Ia-IIIb' possess significant thrombin inhibitory potential.
- The findings contribute to the ongoing debate regarding the genetic versus evolutionary origin of hirudin diversity.