Thromb Haemost 1990; 63(03): 499-504
DOI: 10.1055/s-0038-1645073
Original Article
Schattauer GmbH Stuttgart

Physico-Chemical Properties of Recombinant Desulphatohirudin

A Electricwala
The Division of Biotechnology, Salisbury, England
,
L Irons
1  The Division of Biologies, PHLS Centre for Applied Microbiology & Research, Salisbury, England
,
R Wait
2  The Division of Pathology, PHLS Centre for Applied Microbiology & Research, Salisbury, England
,
R J G Carr
The Division of Biotechnology, Salisbury, England
,
R J Ling
The Division of Biotechnology, Salisbury, England
,
H Grossenbacher
3  The Division of Biotechnology Research Laboratories, CIBA-GEIGY Ltd., Basel, Switzerland
,
T Atkinson
The Division of Biotechnology, Salisbury, England
› Author Affiliations
Further Information

Publication History

Received 26 September 1989

Accepted after revision 23 February 1990

Publication Date:
30 June 2018 (online)

Summary

Physico-chemical properties of recombinant desulphatohirudin expressed in yeast (CIBA GEIGY code No. CGP 39393) were reinvestigated. As previously reported for natural hirudin, the recombinant molecule exhibited abnormal behaviour by gel filtration with an apparent molecular weight greater than that based on the primary structure. However, molecular weight estimation by SDS gel electrophoresis, FAB-mass spectrometry and Photon Correlation Spectroscopy were in agreement with the theoretical molecular weight, with little suggestion of dimer or aggregate formation. Circular dichroism studies of the recombinant molecule show similar spectra at different pH values but are markedly different from that reported by Konno et al. (13) for a natural hirudin-variant. Our CD studies indicate the presence of about 60% beta sheet and the absence of alpha helix in the secondary structure of recombinant hirudin, in agreement with the conformation determined by NMR studies (17)