Ovine platelet factor 4: purification, amino acid sequence, radioimmunoassay and comparison with platelet factor 4 of other species.

A simple method of purification of ovine platelet factor 4 (PF4) is described. Material released by freezing and thawing suspension of washed sheep platelets was fractionated by heparin-agarose chromatography and reverse phase HPLC. Purified ovine PF4 contained 85 amino acids (Mr 9130) and showed 78% homology with bovine PF4, 76% with porcine PF4, 71% homology with human PF4, and 61% with rat PF4. The heparin binding site of ovine PF4 localized in the C-terminal region of the molecule was identified as LYKKIIKRLL. The content of PF4 determined by radioimmunoassay was 22.6 (+/- 1.6 S.D.) micrograms per 10(9) platelets and 46.8 (+/- 19.6 S.D.) ng per ml platelet poor plasma collected in acid citrate dextrose in the presence of prostaglandin E1. PF4 was rapidly released during stimulation of ovine platelets by collagen.