Chromatographie and Electrophoretic Properties of Synthetic Human Fibrinopeptides

Robin McKenzie; D. S Pepper; A. B Kay

doi:10.1055/s-0038-1647733

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1974; 32(02/03): 651-658
DOI: 10.1055/s-0038-1647733

Original Article

Schattauer GmbH

Chromatographie and Electrophoretic Properties of Synthetic Human Fibrinopeptides

Robin McKenzie

¹University Department of Respiratory Diseases, City Hospital, Edinburgh, and the South-East Scotland Regional Blood Transfusion Service, Royal Infirmary of Edinburgh

,

D. S Pepper

¹University Department of Respiratory Diseases, City Hospital, Edinburgh, and the South-East Scotland Regional Blood Transfusion Service, Royal Infirmary of Edinburgh

,

A. B Kay

¹University Department of Respiratory Diseases, City Hospital, Edinburgh, and the South-East Scotland Regional Blood Transfusion Service, Royal Infirmary of Edinburgh

› Author Affiliations

Abstract

Summary

Some properties of synthetic human fibrinopeptides were studied by thin-layer chromatography, thin-layer electrophoresis and low voltage and high voltage paper electrophoresis. The Rf values and electrophoretic mobilities of the peptides in these systems were determined. In high voltage electrophoresis synthetic and natural (fibrinogen-derived) peptides migrated in an identical fashion.

When gel filtration was performed in 0.05 M pyridine or 0.1 N ammonia, synthetic fibrinopeptides A and B appeared to be aggregated. In contrast, when filtration was performed in 1.3 M formic acid, the peptides eluted in positions corresponding to their monomeric molecular weights.

In addition it was possible to quantitate synthetic fibrinopeptides by two colorimetric assays, the Sakaguchi reaction and the Folin-Ciocalteu method. Ultraviolet extinction coefficients for each peptide were also determined.

Full Text

References

References
1 Ambler R. P. 1963; Amino acid sequence of P-Cytochrome-551. Biochemical Journal 89: 352.
2 Blombäck B, Blombäck M. 1956; Purification of human and bovine fibrinogen. Arkiv for Kemi 10: 1956.
3 Blombäck B, Hessel B, Iwanaga S, Reuterby J, Blombäck M. 1972; Primary structure of human fibrinogen and fibrin. I. Cleavage of fibrinogen with cyanogen bromide. Isolation and characterization of NH2-terminal fragments of the a (“A”) chain. Journal of Biological Chemistry 247: 1496.
4 Blombäck B, Vestermark A. 1958; Isolation of fibrino-peptides by chromatography. Arkiv for Kemi 12: 173.
5 Colman R. W, Osbahr A. B, Morris R. E. 1967; New vasoconstrictor, bovine peptide B, released during blood coagulation. Nature 215: 292.
6 Gladner J. A, Murtatjgh P. A, Hoitck J. C. 1968; The biological properties of peptides from fibrinogen. Biochemical Pharmacology. Supplement, 259.
7 Herzig R. H, Ratnoff O. D, Shainoff J. R. 1970; Studies on a procoagulant fraction of southern copperhead snake venom : The preferential release of fibrinopeptide B. Journal of Laboratory and Clinical Medicine 76: 451.
8 Johnson E. J, May W. P. 1969; Rapid peptide synthesis: Synthesis of human fibrinopeptide A. Journal of Pharmaceutical Sciences 58: 1568.
9 Kay A. B, Pepper D. S, McKenzie R. 1974; The identification of fibrinopeptide B as a chemotactic agent derived from human fibrinogen. British Journal of Haematology 27: 669.
10 Leggett-Bailey J. 1967. Techniques in Protein Chemistry. 2nd edn.. Elsevier Publishing Co; Amsterdam: 23-340.
11 Milstein C. P, Milstein C. 1968; A tryptic peptide containing a unique serine phosphate residue in rabbit phosphoglucomutase. Biochemical Journal 109: 93.
12 Shainoff R. S, Page I. H. 1960; Cofibrin and fibrin-intermediates as indicators of thrombin activity in vivo. Circulation Research 08: 1013.